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- Dosage response for compounds that protect hERG from block by proarrhythmic agents using manual patch clamp Data Source: Johns Hopkins Ion Channel Center BioAssay Type: Mode of action, Concentration-Response Relationship Observed Source (MLPCN Center Name): Johns Hopkins Ion Channel Center (JHICC) Center Affiliation: Johns Hopkins University, School of Medicine Screening Center PI: Min Li, Ph.D. Assay Provider: Dr. Sabina Kupershmidt, Vanderbilt University Medical Center Network: Molecular Libraries Probe Production Centers Network (MLPCN) Grant Proposal Number: 1R03MH084820-01 Grant Proposal PI: Dr. Sabina Kupershmidt, Vanderbilt University Medical Center Assay Implementation: Dr. Sabina Kupershmidt, Vanderbilt University Medical Center Name: Dosage response for compounds that protect hERG from block by proarrhythmic agents using manual patch clamp Description: See the related bioassay (AID: 1511). Principle of the assay Patch clamp is gold standard to measure channel activities. Upon depolarization of cells expressing potassium channels, in this case, human ether-a-go-go-related g
- Fluorescence polarization-based biochemical high throughput dose response assay to identify inhibitors that disrupt the binding of a cyclic peptide (Tn6) to the fibrin proteolytic product D-Dimer Source (MLPCN Center Name): The Scripps Research Institute Molecular Screening Center (SRIMSC) Center Affiliation: Harvard Medical School/Massachusetts General Hospital Assay Provider: Peter Caravan, Harvard Medical School/Massachusetts General Hospital Network: Molecular Libraries Probe Production Centers Network (MLPCN) Grant Proposal Number: R21 NS075627 Grant Proposal PI: Peter Caravan, Harvard Medical School/Massachusetts General Hospital External Assay ID: FIBRIN-TN6_INH_FP_1536_3XIC50 DRUN Name: Fluorescence polarization-based biochemical high throughput dose response assay to identify inhibitors that disrupt the binding of a cyclic peptide (Tn6) to the fibrin proteolytic product D-Dimer and fragment E complex [DD(E )]. Description: Fibrin is an insoluble biopolymer that is formed by the proteolytic action of thrombin on the clotting factor fibrinogen. It constitutes the major protein component of blood clots and is generally associated with pathologies such as ischemic stro
- Fluorescence polarization-based biochemical high throughput dose response assay to identify inhibitors that disrupt the binding of a cyclic peptide (Tn7) to the fibrin proteolytic product D-Dimer Source (MLPCN Center Name): The Scripps Research Institute Molecular Screening Center (SRIMSC) Center Affiliation: Harvard Medical School/Massachusetts General Hospital Assay Provider: Peter Caravan, Harvard Medical School/Massachusetts General Hospital Network: Molecular Libraries Probe Production Centers Network (MLPCN) Grant Proposal Number: R21 NS075627 Grant Proposal PI: Peter Caravan, Harvard Medical School/Massachusetts General Hospital External Assay ID: FIBRIN-TN7_INH_FP_1536_3XIC50 DRUN Name: Fluorescence polarization-based biochemical high throughput dose response assay to identify inhibitors that disrupt the binding of a cyclic peptide (Tn7) to the fibrin proteolytic product D-Dimer and fragment E complex [DD(E )]. Description: Fibrin is an insoluble biopolymer that is formed by the proteolytic action of thrombin on the clotting factor fibrinogen. It constitutes the major protein component of blood clots and is generally associated with pathologies such as ischemic stro
- JHICC_CHT_Inh_3H uptake_CRC Data Source (MLPCN Center Name): Johns Hopkins Ion Channel Center (JHICC) Center Affiliation: Johns Hopkins University, School of Medicine Screening Center PI: Min Li, Ph.D. Assay Provider: Alicia Ruggiero, Ph.D., Vanderbilt University Medical Center Network: Molecular Libraries Probe Production Centers Network (MLPCN) Grant Proposal Number: 1R03DA028852-01 Grant Proposal PI: Alicia Ruggiero, Ph.D., Vanderbilt University Medical Center Assay Implementation: Meng Wu Ph.D., Xiaofang Huang, M.S., Zhihong Lin, Ph. D., Kaiping Xu, M.S., Shunyou Long, M.S., and Owen McManus, Ph.D. Description: In the brain, the chemical acetylcholine (ACh) exerts powerful modulatory control over arousal, motor and cognitive circuits, and has been found to be deficient in Alzheimer's Disease (AD). The current drugs available to positively impact cognitive deficits in Alzheimer's Disease (AD) and other dementias are the cholinesterase inhibitors. These prevent the breakdown of the neurotransmitter
- Chemical Antagonists IAP-family anti-apoptotic proteins Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Proposal Number: MH081277-01 Assay Provider: John C. Reed, Sanford-Burnham Medical Research Institute, San Diego, CA Apoptosis plays an essential role in many aspects of normal development and physiology, becoming dysregulated in myriad diseases characterized by insufficient or excessive cell death. Caspases are intracellular proteases that are suppressed by Inhibitor of Apoptosis Proteins (IAPs), a family of evolutionarily conserved anti-apoptotic proteins. Proteins released from mitochondria (SMAC and HtrA2) can competitively displace IAPs from the Caspases, thus helping to drive apoptosis. It has been shown that only a few residues at the N-terminus of activated SMAC protein (4-mer) are sufficient to affect the release of IAPs from Caspases. Thus, it is plausib
- Chemical Antagonists of IAP-family anti-apoptotic proteins confirmation Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Proposal Number: MH081277-01 Assay Provider: John C. Reed, Sanford-Burnham Medical Research Institute, San Diego, CA This XIAP dose response assay is developed and performed to confirm hits originally identified in the XIAP HTS binding assay (AID 1018) and to study the structure-activity relationship on analogs of the confirmed hits. Compounds are either acquired from commercial sources or synthesized internally. Apoptosis plays an essential role in many aspects of normal development and physiology, becoming dysregulated in myriad diseases characterized by insufficient or excessive cell death. Caspases are intracellular proteases that are suppressed by Inhibitor of Apoptosis Proteins (IAPs), a family of evolutionarily conserved anti-apoptotic proteins. Proteins
- Dose Response confirmation of Image-Based HTS for Selective Agonists for NTR1 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Proposal Number: 1 R03 MH089653-01 Assay Provider: Dr. Lawrence Barak, Duke University Medical Center Addiction leading to abuse should be treatable by pharmacological approaches, and programs that identify new drugs to treat methamphetamine abuse address an immediate goal of the National Institute on Drug Abuse (NIDA) that new approaches are needed for treating METH addiction. Currently, small molecule drug-like compounds are not available for treating METH abuse. Neurotensin receptor 1 (NTR1) peptide agonists produce behaviors that are exactly opposite to the psychostimulant effects observed with methamphetamine abuse, such as hyperactivity, neurotoxicity, psychotic episodes, and cognitive deficits, and repeated administrations of NTR1 agonists do not l
- Dose Response confirmation of compounds that inhibit HePTP Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: XO1 MH077603-01 Assay Provider: Dr. Tomas Mustelin, Sanford-Burnham Medical Research Institute Protein tyrosine phosphatases (PTPs), working with protein tyrosine kinases (PTKs), control the phosphorylation state of many proteins in the signal transduction pathways. HePTP is a tyrosine phosphatase expressed in hematopoietic cells and regulates the MAP kinases Erk and p38. It has been found that HePTP is often dysregualted in the preleukemic disorder myelodysplastic syndrome, as well as in acute myelogeneous leukemia. Small molecule inhibitors of HePTP will be useful as molecular probes for studying the mechanism of signal transduction and MAP kinase regulation, and may have therapeutic potential for the treatment of hematopoietic malignancies. Th
- Dose response confirmation of the uHTS fluorescent assay for identification of inhibitors of ATG4B Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH090871-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute, San Diego CA Autophagy is an evolutionarily conserved process whereby cells catabolize damaged proteins and organelles for purposes of generating substrates for sustaining ATP production during times of nutrient deprivation. The autophagic process involves membrane vesicles engulfing cytosol and organelles, delivering their contents to lysosomes for digestion. The genes responsible for autophagy have been identified, largely through genetic analysis of yeast, Saccharomyces cerevisiae, and are conserved in mammals, plants, and essentially all eukaryotes. While autophagy is critical for cell survival in the context of nutrient deprivation, circumstances
- Dose response confirmation of the uHTS fluorescent assay for identification of inhibitors of ATG4B. Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH090871-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute, San Diego CA Autophagy is an evolutionarily conserved process whereby cells catabolize damaged proteins and organelles for purposes of generating substrates for sustaining ATP production during times of nutrient deprivation. The autophagic process involves membrane vesicles engulfing cytosol and organelles, delivering their contents to lysosomes for digestion. The genes responsible for autophagy have been identified, largely through genetic analysis of yeast, Saccharomyces cerevisiae, and are conserved in mammals, plants, and essentially all eukaryotes. While autophagy is critical for cell survival in the context of nutrient deprivation, circumstances
- Fluorescent secondary assay for dose-response confirmation of chemical inhibitors of HePTP Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Number: XO1 MH077603-01 Assay Provider: Dr. Tomas Mustelin, Sanford-Burnham Medical Research Institute Protein tyrosine phosphatases (PTPs), working with protein tyrosine kinases (PTKs), control the phosphorylation state of many proteins in the signal transduction pathways. HePTP is a tyrosine phosphatase expressed in hematopoietic cells and regulates the MAP kinases Erk and p38. It has been found that HePTP is often dysregualted in the preleukemic disorder myelodysplastic syndrome, as well as in acute myelogeneous leukemia. Small molecule inhibitors of HePTP will be useful as molecular probes for studying the mechanism of signal transduction and MAP kinase regulation, and may have therapeutic potential for the treatment of hematopoietic malignancies. This flu
- HTS Discovery of Chemical Inhibitors of HePTP, a Leukemia Target Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Number: XO1 MH077603-01 Assay Provider: Dr. Tomas Mustelin, Sanford-Burnham Medical Research Institute Protein tyrosine phosphatases (PTPs), working with protein tyrosine kinases (PTKs), control the phosphorylation state of many proteins in the signal transduction pathways. HePTP is a tyrosine phosphatase expressed in hematopoietic cells and regulates the MAP kinases Erk and p38. It has been found that HePTP is often dysregualted in the preleukemic disorder myelodysplastic syndrome, as well as in acute myelogeneous leukemia. Small molecule inhibitors of HePTP will be useful as molecular probes for studying the mechanism of signal transduction and MAP kinase regulation, and may have therapeutic potential for the treatment of hematopoietic malignancies. This colorim
- Image-based HTS for Selective Agonists of GPR55 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Proposal Number: 1X01 DA026205-01 Assay Provider: Dr. Mary Abood, California Pacific Medical Center Research Institute (currently Temple University) The cannabinoid and endocannabinoid system has been implicated in the pathophysiology of drug dependence and addiction disorders. GPR55, an orphan G-Protein Coupled Receptor, has been reported to be a cannabinoid receptor, but its status as such remains unresolved due to conflicting results from pharmacological studies. The goal of the project is to identify small molecule agonists of GPR55, which may aid in the deorphanization efforts of this receptor and ultimately further the understanding of the role of GPR55 in drug addiction. This high content imaging assay utilizes a cell line permanently expressing a
- In Vitro Hsc70 Dose Response Fluorescence Polarization Assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Screening Centers Network (MLSCN) MLSCN Grant: XO1 MH079863-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute ) This Hsc70 dose response assay is developed and performed to study the specificity of analogs of hits tested in the "In Vitro Hsp70 Dose Response Fluorescence Polarization Assay for SAR Study" (AID 1072). Over-expression of molecular chaperones occurs commonly in cancers and provides protection from a wide variety of cellular stresses, both endogenous and iatrogenic. Molecular chaperones also play important roles in maintaining the activity of several signal-transducing proteins and transcriptions factors involved in malignant transformation. The human genome contains nine Hsp70-family genes. These chaperones include Hsp70 and Hsc70, which are commonly over-ex
- QFRET-based biochemical high throughput dose response assay for inhibitors of the Plasmodium falciparum M18 Aspartyl Aminopeptidase (PFM18AAP). Source (MLPCN Center Name): The Scripps Research Institute Molecular Screening Center Affiliation: The Scripps Research Institute, TSRI Assay Provider: John Dalton and Donald Gardiner, Queensland Institute of Medical Research, Australia Network: Molecular Library Probe Production Centers Network (MLPCN) Grant Proposal Number: 1 R03 MH084103-01 Grant Proposal PI: John Dalton and Donald Gardiner, Queensland Institute of Medical Research, Australia External Assay ID: PFM18AAP_INH_QFRET_1536_3XIC50 Name: QFRET-based biochemical high throughput dose response assay for inhibitors of the Plasmodium falciparum M18 Aspartyl Aminopeptidase (PFM18AAP). Description: Aminopeptidases (APs) are metalloproteases that cleave amino-terminal (N-terminal) amino acids during protein synthesis (1, 2) These enzymes are characterized in part by their post-translational removal of leucine, aspartate, proline, methionine, etc from proteins and peptides, in order that proteins are properly regulated, targete
- SAR analysis of Antagonists of IAP-family anti-apoptotic proteins Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Proposal Number: MH081277-01 Assay Provider: John C. Reed, Sanford-Burnham Medical Research Institute, San Diego, CA This XIAP dose response assay is developed and performed to confirm hits originally identified in the XIAP HTS binding assay (AID 1018) and to study the structure-activity relationship on analogs of the confirmed hits. Compounds are either acquired from commercial sources or synthesized internally. The assay was performed in the assay providers' laboratory. Apoptosis plays an essential role in many aspects of normal development and physiology, becoming dysregulated in myriad diseases characterized by insufficient or excessive cell death. Caspases are intracellular proteases that are suppressed by Inhibitor of Apoptosis Proteins (IAPs), a fami
- SAR analysis of an In Vitro TNAP Dose Response Luminescent Assay Sanford-Burnham Center for Chemical Genomics (SBCCG) Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Number: MH077602-01 Assay Provider: Dr. Jose Luis Millan, Sanford-Burnham Medical Research Institute, San Diego, CA This TNAP dose response assay is developed and performed for the purpose of SAR study on analogs of hits originally identified in the TNAP luminescent HTS assay (AID 518) Alkaline phosphatase (EC 3.1.3.1) (APs) catalyze the hydrolysis of phosphomonoesters, releasing phosphate and alcohol. APs are dimeric enzymes found in the most organism. In human, four isozymes of APs have been identified. Three isozymes are tissue-specific and the fourth one is tissue-nonsepecifc, named TNAP. TNAP deficiency is associated with defective bone mineralization in the form of rickets and osteomalacia. Therefore, there are therapeutic potentials of inhibiting TNAP activity
- SAR analysis of compounds that inhibit HePTP - Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLSCN) Grant Number: XO1 MH077603-01 Assay Provider: Dr. Tomas Mustelin, Sanford-Burnham Medical Research Institute Protein tyrosine phosphatases (PTPs), working with protein tyrosine kinases (PTKs), control the phosphorylation state of many proteins in the signal transduction pathways. HePTP is a tyrosine phosphatase expressed in hematopoietic cells and regulates the MAP kinases Erk and p38. It has been found that HePTP is often dysregualted in the preleukemic disorder myelodysplastic syndrome, as well as in acute myelogeneous leukemia. Small molecule inhibitors of HePTP will be useful as molecular probes for studying the mechanism of signal transduction and MAP kinase regulation, and may have therapeutic potential for the treatment of hematopoietic malignancies. Th
- SAR analysis of compounds that potentiate TRAIL-induced apoptosis in PPC-1 cells. Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: X01 MH083230-01 Assay Provider: Dr. Dmitri Rozanov, Sanford-Burnham Medical Research Institute, San Diego CA This dose response assay is developed and performed to confirm hits originally identified in "uHTS for the identification of compounds that potentiate TRAIL-induced apoptosis of cancer cells" (AID 1443) and to study the structure-activity relationship on analogs of the confirmed hits. Compounds are either acquired from commercial sources or synthesized internally. Cytotoxic chemotherapy induces apoptosis via a pathway involving mitochondria, sometimes referred to as the "intrinsic pathway." An acquired resistance to anticancer drugs commonly results from the accumulation of defects in components of the mitochondrial pathway for apoptosis. Discov
- Colorimetric assay for HTS discovery of chemical inhibitors of EphA4 receptor antagonists Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Number: XO1 MH077603-01 Assay Provider: Dr. Elena Pasquale, Sanford-Burnham Medical Research Institute EphA4 is a member of the large Eph family of receptor tyrosine kinases. The signaling ability of EphA4, and the other nine closely related EphA receptors, is activated by binding the six GPI-linked ephrin-A ligands. In addition, EphA4 also binds the three transmembrane ephrin-B ligands, which are the ligands for the other class of Eph receptors, the EphB receptors (EphB1-EphB6). Eph receptor-ephrin interaction requires cell-cell contact because both the receptor and the ligand are membrane-bound. Importantly, signals are generated both through the Eph receptor kinase domain (forward signals) and through signaling molecules associated with the ephrins (reverse
- Counter Screen using XIAP-Bir3 of the Chemical Antagonists of IAP-family anti-apoptotic proteins confirmation assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Proposal Number: MH081277-01 Assay Provider: John C. Reed, Sanford-Burnham Medical Research Institute, San Diego, CA This dose response assay is developed and performed as a counter screen to compounds in the Chemical Antagonists of IAP-family anti-apoptotic proteins confirmation (AID 1449) and to study the structure-activity relationship on analogs of the confirmed hits. Compounds are either acquired from commercial sources or synthesized internally. Apoptosis plays an essential role in many aspects of normal development and physiology, becoming dysregulated in myriad diseases characterized by insufficient or excessive cell death. Caspases are intracellular proteases that are suppressed by Inhibitor of Apoptosis Proteins (IAPs), a family of evolutionarily conse
- Counter screen SAR assay for PMM2 inhibitors via a fluorescence intensity assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Probe Production Centers Network (MLPCN) Grant Number: R03 MH082386-01 Assay Provider: Dr. Hudson H. Freeze, Sanford-Burnham Medical Research Institute, San Diego, CA Congenital Disorders of Glycosylation (CDGs) are rare genetic disorders in the synthesis of N-linked glycan chains. Mutations in PMM2, encoding phosphomannomutase 2 (PMM2, Man-6-P-> Man-1-P) cause the most common form, CDG-Ia. Patients have a host of problems including hypotonia, variable psychomotor retardation, seizures, peripheral neuropathy, cardiomyopathy, and protein losing enteropathy. There is no therapy for this disorder. A current approach to ameliorate the physiological conditions associated with CDG-Ia is to provide high influx of mannose for patience. We previously developed a HTS assay through the MLSCN to identify inhibitors of phos
- Counterscreen for inhibitors of M1 and M17 aminopeptidases: QFRET-based biochemical high throughput dose response assay for inhibitors of the Cathepsin L proteinase (CTSL1). Source (MLPCN Center Name): The Scripps Research Institute Molecular Screening Center Affiliation: The Scripps Research Institute, TSRI Assay Provider: John Dalton and Donald Gardiner, Queensland Institute of Medical Research, Australia Network: Molecular Library Probe Production Centers Network (MLPCN) Grant Proposal Number: 1 R03 MH084103-01 Grant Proposal PI: John Dalton and Donald Gardiner, Queensland Institute of Medical Research, Australia External Assay ID: CTSL1_INH_QFRET_1536_3XIC50 CSDRUN Name: Counterscreen for inhibitors of M1 and M17 aminopeptidases: QFRET-based biochemical high throughput dose response assay for inhibitors of the Cathepsin L proteinase (CTSL1). Description: Aminopeptidases (APs) are metalloproteases that cleave amino-terminal (N-terminal) amino acids during protein synthesis (1, 2) These enzymes are characterized in part by their post-translational removal of leucine, aspartate, proline, methionine, etc from proteins and peptides, in order that protei
- Dose Response Confirmation of compounds that inhibit VHR1 in Fluorescent Assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH084230-01A1 Assay Provider: Dr. Lutz Tautz, Source Affiliation: Sanford-Burnham Medical Research Institute (San Diego, CA) Protein tyrosine phosphatases (PTPs) play vital roles in numerous cellular processes and are implicated in a growing number of human diseases, ranging from cancer to cardiovascular, immunological, infectious, neurological, and metabolic diseases. The Vaccinia H1-related (VHR) PTP is a dual-specific Erk and Jnk phosphatase, the loss of which causes specific cell cycle arrest in HeLa carcinoma cells, suggesting that VHR inhibition may be a useful approach to halt the growth of cancer cells without detrimental effects on normal cells. Recent studies by collaborators and us suggest that VHR is upregulated in several cervix
- Dose Response concentration confirmation of uHTS hits from a small molecule activators of human intestinal alkaline phosphatase via a luminescent assay - Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Proposal Number: X01-MH077602-01 Assay Provider Dr. Jose Luis Millan, Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Alkaline phosphatase (EC 3.1.3.1) (APs) catalyze the hydrolysis of phosphomonoesters, releasing inorganic phosphate and alcohol. APs are dimeric enzymes found in most organisms. In human, four isozymes of APs have been identified. One isozyme is tissue-nonspecific (designated TNAP) and three other isozymes are tissue-specific and named according to the tissue of their predominant expression: intestinal (IAP), placental (PLAP) and germ cell (GCAP) alkaline phosphatases. IAP expression is largely restricted to the gut, especially to the epithelial cells (enterocytes) of the small intestinal mucosa. The exact biological function of
- Dose Response concentration confirmation of uHTS hits from a small molecule activators of human intestinal alkaline phosphatase via a luminescent assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Proposal Number: X01-MH077602-01 Assay Provider Dr. Jose Luis Millan, Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Alkaline phosphatase (EC 3.1.3.1) (APs) catalyze the hydrolysis of phosphomonoesters, releasing inorganic phosphate and alcohol. APs are dimeric enzymes found in most organisms. In human, four isozymes of APs have been identified. One isozyme is tissue-nonspecific (designated TNAP) and three other isozymes are tissue-specific and named according to the tissue of their predominant expression: intestinal (IAP), placental (PLAP) and germ cell (GCAP) alkaline phosphatases. IAP expression is largely restricted to the gut, especially to the epithelial cells (enterocytes) of the small intestinal mucosa. The exact biological function of
- Dose Response confirmation of HTS hits from an HePTP Fluorescent Assay using OMFP substrate - Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: XO1 MH077603-01 Assay Provider: Dr. Tomas Mustelin, Sanford-Burnham Medical Research Institute Protein tyrosine phosphatases (PTPs), working with protein tyrosine kinases (PTKs), control the phosphorylation state of many proteins in the signal transduction pathways. HePTP is a tyrosine phosphatase expressed in hematopoietic cells and regulates the MAP kinases Erk and p38. It has been found that HePTP is often dysregualted in the preleukemic disorder myelodysplastic syndrome, as well as in acute myelogeneous leukemia. Small molecule inhibitors of HePTP will be useful as molecular probes for studying the mechanism of signal transduction and MAP kinase regulation, and may have therapeutic potential for the treatment of hematopoietic malignancies. Th
- Dose Response confirmation of UBC13 Polyubiquitin Inhibitors using a Bfl-1 counterscreen Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1R03 MH085677-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute, San Diego CA Tumor Necrosis Factor Receptor-Associated Factors (TRAFs) are a family of adapter proteins that bind an unusual ubiquitin-conjugating enzyme, Ubc13, which produces polyubiquitin chains linked at lysine 63 of ubiquitin. These lysine 63-linked ubiquitin polymers trigger changes in protein activity. Ubiquitination by Ubc13 of TRAFs and the various protein kinases to which TRAFs bind is recognized as a critical step in signaling by TNFRs, TLRs, NLRs, and T-cell and B-cell antigen receptors (TCR/BCR) during innate and acquired immune responses. Since aberrant signaling by these receptor systems is linked to a wide variety of autoimmune, inflammato
- Dose Response confirmation of inhibitors of NALP1 in yeast using a Caspase-1-ASC counter screen. Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1 U01 AI078048 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute, San Diego, CA NLR family proteins are an important component of the innate immune system of vertebrates. These proteins possess a nucleotide-binding oligomerization domain, called NACHT, in combination with variable numbers of Leucine-Rich Repeat (LRR) domains that bind molecules produced by pathogens and probably also products of tissue injury. Among the effector mechanisms of NLR family proteins is activation of Caspase-1, which cleaves and activates pro-inflammatory cytokines. We present here a unique primary assay, in which we have reconstituted the mammalian Caspase mediated IL-1 activation pathway consisting of NLRP1 (NALP1), ASC, and Caspase-1 in Sacchar
- Dose Response confirmation of inhibitors of NALP3 in yeast using a Caspase-1-ASC counter screen Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1 U01 AI078048 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute, San Diego CA NLR family proteins are an important component of the innate immune system of vertebrates. These proteins possess a nucleotide-binding oligomerization domain, called NACHT, in combination with variable numbers of Leucine-Rich Repeat (LRR) domains that bind molecules produced by pathogens and probably also products of tissue injury. Among the effector mechanisms of NLR family proteins is activation of Caspase-1, which cleaves and activates pro-inflammatory cytokines. We present here a unique primary assay, in which we have reconstituted the mammalian Caspase mediated IL-1 activation pathway consisting of NLRP3 (NALP3), ASC, and Caspase-1 in Saccharo
- Dose Response confirmation of inhibitors of Sentrin-specific proteases (SENPs) using a Caspase-3 Selectivity assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network(MLPCN) Grant Proposal Number: 1R21 NS061758-01 fast track Assay Provider: Dr. Guy Salvesen, Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Modification of proteins by SUMO is a dynamic and reversible process. SUMOylation/deSUMOylation cycle regulates SUMOs function. Sentrin-specific proteases (SENPs) are involved in both the maturation of SUMO precursors (endopeptidase cleavage) and deconjugation of the targets (isopeptidase cleavage) [1-3]. There are seven SENPs (1, 2, 3, 5, 6, 7, 8) in humans, and several of these have been characterized as SUMO (or Nedd8) specific enzymes. SENP8 is not a SUMO protease, instead it functions on a small ubiquitin related protein Nedd8. The objective of this project is to generate small molecule inhibitors specific for
- Dose Response confirmation of inhibitors of Sentrin-specific proteases (SENPs) using a Luminescent Interference Counterscreen assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network(MLPCN)Grant Proposal Number: 1R21 NS061758-01 fast track Assay Provider: Dr. Guy Salvesen, Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Modification of proteins by SUMO is a dynamic and reversible process. SUMOylation/deSUMOylation cycle regulates SUMOs function. Sentrin-specific proteases (SENPs) are involved in both the maturation of SUMO precursors (endopeptidase cleavage) and deconjugation of the targets (isopeptidase cleavage) [1-3]. There are seven SENPs (1, 2, 3, 5, 6, 7, 8) in humans, and several of these have been characterized as SUMO (or Nedd8) specific enzymes. SENP8 is not a SUMO protease, instead it functions on a small ubiquitin related protein Nedd8. The objective of this project is to generate small molecule inhibitors specific for SENP8
- Dose Response confirmation of small molecule antagonists of the CCR6 receptor: a luminescent beta-arrestin assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: R21 NS064746-01A Assay Provider: Dr. Greg Roth, Sanford-Burnham Medical Research Institute Currently there are no published patents or studies specifically describing small molecule antagonists of the chemokine receptor CCR6. CCL20 (MIP-3 alpha) is the endogenous peptide ligand for the G-protein coupled receptor (GPCR) CCR6. The receptor ligand pair is responsible for the chemoattraction of immature dendritic cells, effector/memory T cells, B cells, and also plays a role at skin and mucosal surfaces. The CCR6 receptor is expressed by B cells, subsets of T cells, and dendritic cells (DC). The link between the CCR6lCCL20 axis and cancer cell metastasis is a recent finding. There are two key studies that describe a relation between CCR6 and colorectal
- Dose Response confirmation of uHTS activators of Human Intestinal Alkaline Phosphatase using Placental Alkaline Phosphatase Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Proposal Number: X01-MH077602-01 Assay Provider Dr. Jose Luis Millan, Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Alkaline phosphatase (EC 3.1.3.1) (APs) catalyze the hydrolysis of phosphomonoesters, releasing inorganic phosphate and alcohol. APs are dimeric enzymes found in most organisms. In human, four isozymes of APs have been identified. One isozyme is tissue-nonspecific (designated TNAP) and three other isozymes are tissue-specific and named according to the tissue of their predominant expression: intestinal (IAP), placental (PLAP) and germ cell (GCAP) alkaline phosphatases. IAP expression is largely restricted to the gut, especially to the epithelial cells (enterocytes) of the small intestinal mucosa. The exact biological function of
- Dose Response confirmation of uHTS activators of Human Intestinal Alkaline Phosphatase using Tissue Nonspecific Alkaline Phosphatase. Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Proposal Number: X01-MH077602-01 Assay Provider Dr. Jose Luis Millan, Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Alkaline phosphatase (EC 3.1.3.1) (APs) catalyze the hydrolysis of phosphomonoesters, releasing inorganic phosphate and alcohol. APs are dimeric enzymes found in most organisms. In human, four isozymes of APs have been identified. One isozyme is tissue-nonspecific (designated TNAP) and three other isozymes are tissue-specific and named according to the tissue of their predominant expression: intestinal (IAP), placental (PLAP) and germ cell (GCAP) alkaline phosphatases. IAP expression is largely restricted to the gut, especially to the epithelial cells (enterocytes) of the small intestinal mucosa. The exact biological function of
- Dose Response confirmation of uHTS activators of Mouse Intestinal Alkaline Phosphatase using Human Intestinal Alkaline Phosphatase Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Proposal Number: X01-MH077602-01 Assay Provider Dr. Jose Luis Millan, Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Alkaline phosphatase (EC 3.1.3.1) (APs) catalyze the hydrolysis of phosphomonoesters, releasing inorganic phosphate and alcohol. APs are dimeric enzymes found in most organisms. In human, four isozymes of APs have been identified. One isozyme is tissue-nonspecific (designated TNAP) and three other isozymes are tissue-specific and named according to the tissue of their predominant expression: intestinal (IAP), placental (PLAP) and germ cell (GCAP) alkaline phosphatases. IAP expression is largely restricted to the gut, especially to the epithelial cells (enterocytes) of the small intestinal mucosa. The exact biological function of
- Dose Response confirmation of uHTS for inhibitors of Sentrin-specific protease 6 (SENP6) using a Luminescent assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network(MLPCN) Grant Proposal Number: 1R21 NS061758-01 fast track Assay Provider: Dr. Guy Salvesen, Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Modification of proteins by SUMO is a dynamic and reversible process. SUMOylation/deSUMOylation cycle regulates SUMOs function. Sentrin-specific proteases (SENPs) are involved in both the maturation of SUMO precursors (endopeptidase cleavage) and deconjugation of the targets (isopeptidase cleavage) [1-3]. There are seven SENPs (1, 2, 3, 5, 6, 7, 8) in humans, and several of these have been characterized as SUMO (or Nedd8) specific enzymes. The objective of this project is to generate small molecule inhibitors specific for SENP6 (the deSUMOylating enzyme). 1536-well chemiluminescent screening assay utilizes RLRGG-
- Dose Response confirmation of uHTS for inhibitors of Sentrin-specific protease 8 (SENP8) using a Luminescent assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network(MLPCN)Grant Proposal Number: 1R21 NS061758-01 fast track Assay Provider: Dr. Guy Salvesen, Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Modification of proteins by SUMO is a dynamic and reversible process. SUMOylation/deSUMOylation cycle regulates SUMOs function. Sentrin-specific proteases (SENPs) are involved in both the maturation of SUMO precursors (endopeptidase cleavage) and deconjugation of the targets (isopeptidase cleavage) [1-3]. There are seven SENPs (1, 2, 3, 5, 6, 7, 8) in humans, and several of these have been characterized as SUMO (or Nedd8) specific enzymes. SENP8 is not a SUMO protease, instead it functions on a small ubiquitin related protein Nedd8. The objective of this project is to generate small molecule inhibitors specific for SENP8
- Dose Response confirmation of uHTS for the identification of UBC13 Polyubiquitin Inhibitors via a TR-FRET Assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1R03 MH085677-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute, San Diego CA Tumor Necrosis Factor Receptor-Associated Factors (TRAFs) are a family of adapter proteins that bind an unusual ubiquitin-conjugating enzyme, Ubc13, which produces polyubiquitin chains linked at lysine 63 of ubiquitin. These lysine 63-linked ubiquitin polymers trigger changes in protein activity. Ubiquitination by Ubc13 of TRAFs and the various protein kinases to which TRAFs bind is recognized as a critical step in signaling by TNFRs, TLRs, NLRs, and T-cell and B-cell antigen receptors (TCR/BCR) during innate and acquired immune responses. Since aberrant signaling by these receptor systems is linked to a wide variety of autoimmune, inflammato
- Dose Response confirmation of uHTS for the identification of UBC13 Polyubiquitin Inhibitors via a TR-FRET Assay reconfirm Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1R03 MH085677-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute, San Diego CA Tumor Necrosis Factor Receptor-Associated Factors (TRAFs) are a family of adapter proteins that bind an unusual ubiquitin-conjugating enzyme, Ubc13, which produces polyubiquitin chains linked at lysine 63 of ubiquitin. These lysine 63-linked ubiquitin polymers trigger changes in protein activity. Ubiquitination by Ubc13 of TRAFs and the various protein kinases to which TRAFs bind is recognized as a critical step in signaling by TNFRs, TLRs, NLRs, and T-cell and B-cell antigen receptors (TCR/BCR) during innate and acquired immune responses. Since aberrant signaling by these receptor systems is linked to a wide variety of autoimmune, inflammato
- Dose Response confirmation of uHTS for the identification of inhibitors of NALP1 in yeast using a luminescent assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1 U01 AI078048 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute, San Diego, CA NLR family proteins are an important component of the innate immune system of vertebrates. These proteins possess a nucleotide-binding oligomerization domain, called NACHT, in combination with variable numbers of Leucine-Rich Repeat (LRR) domains that bind molecules produced by pathogens and probably also products of tissue injury. Among the effector mechanisms of NLR family proteins is activation of Caspase-1, which cleaves and activates pro-inflammatory cytokines. We present here a unique primary assay, in which we have reconstituted the mammalian Caspase mediated IL-1 activation pathway consisting of NLRP1 (NALP1), ASC, and Caspase-1 in Sacchar
- Dose Response confirmation of uHTS for the identification of inhibitors of NALP3 in yeast using a luminescent assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1 U01 AI078048 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute, San Diego CA NLR family proteins are an important component of the innate immune system of vertebrates. These proteins possess a nucleotide-binding oligomerization domain, called NACHT, in combination with variable numbers of Leucine-Rich Repeat (LRR) domains that bind molecules produced by pathogens and probably also products of tissue injury. Among the effector mechanisms of NLR family proteins is activation of Caspase-1, which cleaves and activates pro-inflammatory cytokines. We present here a unique primary assay, in which we have reconstituted the mammalian Caspase mediated IL-1 activation pathway consisting of NLRP3 (NALP3), ASC, and Caspase-1 in Saccharo
- Dose Response confirmation of uHTS hits from a small molecule inhibitors of human intestinal alkaline phosphatase via a luminescent assay - Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Proposal Number: X01-MH077602-01 Assay Provider Dr. Jose Luis Millan, Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Alkaline phosphatase (EC 3.1.3.1) (APs) catalyze the hydrolysis of phosphomonoesters, releasing inorganic phosphate and alcohol. APs are dimeric enzymes found in most organisms. In human, four isozymes of APs have been identified. One isozyme is tissue-nonspecific (designated TNAP) and three other isozymes are tissue-specific and named according to the tissue of their predominant expression: intestinal (IAP), placental (PLAP) and germ cell (GCAP) alkaline phosphatases. IAP expression is largely restricted to the gut, especially to the epithelial cells (enterocytes) of the small intestinal mucosa. The exact biological functio
- Dose Response confirmation of uHTS hits from a small molecule inhibitors of human intestinal alkaline phosphatase via a luminescent assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Proposal Number: X01-MH077602-01 Assay Provider Dr. Jose Luis Millan, Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Alkaline phosphatase (EC 3.1.3.1) (APs) catalyze the hydrolysis of phosphomonoesters, releasing inorganic phosphate and alcohol. APs are dimeric enzymes found in most organisms. In human, four isozymes of APs have been identified. One isozyme is tissue-nonspecific (designated TNAP) and three other isozymes are tissue-specific and named according to the tissue of their predominant expression: intestinal (IAP), placental (PLAP) and germ cell (GCAP) alkaline phosphatases. IAP expression is largely restricted to the gut, especially to the epithelial cells (enterocytes) of the small intestinal mucosa. The exact biological function of
- Dose Response confirmation of uHTS hits from a small molecule inhibitors of mouse intestinal alkaline phosphatase via a luminescent assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Proposal Number: X01-MH077602-01 Assay Provider Dr. Jose Luis Millan, Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Alkaline phosphatase (EC 3.1.3.1) (APs) catalyze the hydrolysis of phosphomonoesters, releasing inorganic phosphate and alcohol. APs are dimeric enzymes found in most organisms. In human, four isozymes of APs have been identified. One isozyme is tissue-nonspecific (designated TNAP) and three other isozymes are tissue-specific and named according to the tissue of their predominant expression: intestinal (IAP), placental (PLAP) and germ cell (GCAP) alkaline phosphatases. IAP expression is largely restricted to the gut, especially to the epithelial cells (enterocytes) of the small intestinal mucosa. The exact biological function of
- Dose Response confirmation of uHTS inhibitors of Human Intestinal Alkaline Phosphatase using Mouse Intestinal Alkaline Phosphatase Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Proposal Number: X01-MH077602-01 Assay Provider Dr. Jose Luis Millan, Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Alkaline phosphatase (EC 3.1.3.1) (APs) catalyze the hydrolysis of phosphomonoesters, releasing inorganic phosphate and alcohol. APs are dimeric enzymes found in most organisms. In human, four isozymes of APs have been identified. One isozyme is tissue-nonspecific (designated TNAP) and three other isozymes are tissue-specific and named according to the tissue of their predominant expression: intestinal (IAP), placental (PLAP) and germ cell (GCAP) alkaline phosphatases. IAP expression is largely restricted to the gut, especially to the epithelial cells (enterocytes) of the small intestinal mucosa. The exact biological function of
- Dose Response confirmation of uHTS inhibitors of Human Intestinal Alkaline Phosphatase using Placental Alkaline Phosphatase Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Proposal Number: X01-MH077602-01 Assay Provider Dr. Jose Luis Millan, Sanford-Burnham Medical Research Institute, San Diego, CA) Alkaline phosphatase (EC 3.1.3.1) (APs) catalyze the hydrolysis of phosphomonoesters, releasing inorganic phosphate and alcohol. APs are dimeric enzymes found in most organisms. In human, four isozymes of APs have been identified. One isozyme is tissue-nonspecific (designated TNAP) and three other isozymes are tissue-specific and named according to the tissue of their predominant expression: intestinal (IAP), placental (PLAP) and germ cell (GCAP) alkaline phosphatases. IAP expression is largely restricted to the gut, especially to the epithelial cells (enterocytes) of the small intestinal mucosa. The exact biological function of I
- Dose Response confirmation of uHTS inhibitors of Human Intestinal Alkaline Phosphatase using Tissue Nonspecific Alkaline Phosphatase. Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Proposal Number: X01-MH077602-01 Assay Provider Dr. Jose Luis Millan, Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Alkaline phosphatase (EC 3.1.3.1) (APs) catalyze the hydrolysis of phosphomonoesters, releasing inorganic phosphate and alcohol. APs are dimeric enzymes found in most organisms. In human, four isozymes of APs have been identified. One isozyme is tissue-nonspecific (designated TNAP) and three other isozymes are tissue-specific and named according to the tissue of their predominant expression: intestinal (IAP), placental (PLAP) and germ cell (GCAP) alkaline phosphatases. IAP expression is largely restricted to the gut, especially to the epithelial cells (enterocytes) of the small intestinal mucosa. The exact biological functio
- Dose Response confirmation of uHTS inhibitors of Mouse Intestinal Alkaline Phosphatase using Human Intestinal Alkaline Phosphatase Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Proposal Number: X01-MH077602-01 Assay Provider Dr. Jose Luis Millan, Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Alkaline phosphatase (EC 3.1.3.1) (APs) catalyze the hydrolysis of phosphomonoesters, releasing inorganic phosphate and alcohol. APs are dimeric enzymes found in most organisms. In human, four isozymes of APs have been identified. One isozyme is tissue-nonspecific (designated TNAP) and three other isozymes are tissue-specific and named according to the tissue of their predominant expression: intestinal (IAP), placental (PLAP) and germ cell (GCAP) alkaline phosphatases. IAP expression is largely restricted to the gut, especially to the epithelial cells (enterocytes) of the small intestinal mucosa. The exact biological functio
- Dose Response confirmation of uHTS inhibitors of Mouse Intestinal Alkaline Phosphatase using Placental Alkaline Phosphatase Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Proposal Number: X01-MH077602-01 Assay Provider Dr. Jose Luis Millan, Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Alkaline phosphatase (EC 3.1.3.1) (APs) catalyze the hydrolysis of phosphomonoesters, releasing inorganic phosphate and alcohol. APs are dimeric enzymes found in most organisms. In human, four isozymes of APs have been identified. One isozyme is tissue-nonspecific (designated TNAP) and three other isozymes are tissue-specific and named according to the tissue of their predominant expression: intestinal (IAP), placental (PLAP) and germ cell (GCAP) alkaline phosphatases. IAP expression is largely restricted to the gut, especially to the epithelial cells (enterocytes) of the small intestinal mucosa. The exact biological function of
- Dose Response confirmation of uHTS inhibitors of Mouse Intestinal Alkaline Phosphatase using Tissue Nonspecific Alkaline Phosphatase. Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Proposal Number: X01-MH077602-01 Assay Provider Dr. Jose Luis Millan, Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Alkaline phosphatase (EC 3.1.3.1) (APs) catalyze the hydrolysis of phosphomonoesters, releasing inorganic phosphate and alcohol. APs are dimeric enzymes found in most organisms. In human, four isozymes of APs have been identified. One isozyme is tissue-nonspecific (designated TNAP) and three other isozymes are tissue-specific and named according to the tissue of their predominant expression: intestinal (IAP), placental (PLAP) and germ cell (GCAP) alkaline phosphatases. IAP expression is largely restricted to the gut, especially to the epithelial cells (enterocytes) of the small intestinal mucosa. The exact biological function of
- Dose Response selectivity of inhibitors of STriatal-Enriched Phosphatase (STEP) in the Lymphoid Phosphatase (PTPN22) Inhibition Assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1R03MH095532-01 Assay Provider: Dr. Lutz Tautz, Sanford-Burnham Medical Research Institute, San Diego CA Disturbance of the dynamic balance between protein tyrosine phosphorylation and dephosphorylation is crucial for the development of many serious conditions, including cancer, diabetes, and autoimmune disorders. This is the first time that tyrosine phosphatase inhibitors are being proposed to improve cognitive function in Alzheimer's disease (AD). STriatal-Enriched Phosphatase (STEP) is a brain-specific protein tyrosine phosphatase that is highly expressed in regions where consolidation of memory occurs and regulates the internalization of NMDARs. Our recent work demonstrates that STEP is elevated in the prefrontal cortex of human AD patients
- Dose Response selectivity of inhibitors of STriatal-Enriched Phosphatase (STEP) in the dual-specificity protein-tyrosine phosphatase VHR Inhibition Assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1R03MH095532-01 Assay Provider: Dr. Lutz Tautz, Sanford-Burnham Medical Research Institute, San Diego CA Disturbance of the dynamic balance between protein tyrosine phosphorylation and dephosphorylation is crucial for the development of many serious conditions, including cancer, diabetes, and autoimmune disorders. This is the first time that tyrosine phosphatase inhibitors are being proposed to improve cognitive function in Alzheimer's disease (AD). STriatal-Enriched Phosphatase (STEP) is a brain-specific protein tyrosine phosphatase that is highly expressed in regions where consolidation of memory occurs and regulates the internalization of NMDARs. Our recent work demonstrates that STEP is elevated in the prefrontal cortex of human AD patients
- Dose Response selectivity of inhibitors of Striatal-Enriched Phosphatase (STEP) in a SHP2 (PTPN11) Inhibition Assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1R03MH095532-01 Assay Provider: Dr. Lutz Tautz, Sanford-Burnham Medical Research Institute, San Diego CA Disturbance of the dynamic balance between protein tyrosine phosphorylation and dephosphorylation is crucial for the development of many serious conditions, including cancer, diabetes, and autoimmune disorders. This is the first time that tyrosine phosphatase inhibitors are being proposed to improve cognitive function in Alzheimer's disease (AD). STriatal-Enriched Phosphatase (STEP) is a brain-specific protein tyrosine phosphatase that is highly expressed in regions where consolidation of memory occurs and regulates the internalization of NMDARs. Our recent work demonstrates that STEP is elevated in the prefrontal cortex of human AD patients
- Dose response confirmation of uHTS small molecule inhibitors of Striatal-Enriched Phosphatase via a fluorescence intensity assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1R03MH095532-01 Assay Provider: Dr. Lutz Tautz, Sanford-Burnham Medical Research Institute, San Diego CA Disturbance of the dynamic balance between protein tyrosine phosphorylation and dephosphorylation is crucial for the development of many serious conditions, including cancer, diabetes, and autoimmune disorders. This is the first time that tyrosine phosphatase inhibitors are being proposed to improve cognitive function in Alzheimer's disease (AD). STriatal-Enriched Phosphatase (STEP) is a brain-specific protein tyrosine phosphatase that is highly expressed in regions where consolidation of memory occurs and regulates the internalization of NMDARs. Our recent work demonstrates that STEP is elevated in the prefrontal cortex of human AD patients
- Dose response counterscreen of uHTS hits for ATG4B inhibitors in a Phospholipase A2 assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH090871-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute, San Diego CA This assay is a counterscreen for the primary screen (AID504462) which looked for inhibitors of Autophagin 1 (ATG4B). In screening for compounds that inhibit ATG4B, the High Throughput Screening (HTS) assay utilized a cleavable form of Phospholipase A2 (PLA2), which is expressed as a fusion protein with the Autophagin substrate LC3/ATG8 appended to its N-terminus, as the substrate for the primary enzymatic reaction. The addition of sequences to the N-terminus of PLA2 inhibits the activity of this enzyme. Cleavage by proteases removing the N-terminal extension then restores enzyme activity, constituting the basis for a protease assay. The sub
- Dose response counterscreen of uHTS hits for ATG4B inhibitors in a Phospholipase A2 assay Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH090871-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute, San Diego CA This assay is a counterscreen for the primary screen (AID504462) which looked for inhibitors of Autophagin 1 (ATG4B). In screening for compounds that inhibit ATG4B, the High Throughput Screening (HTS) assay utilized a cleavable form of Phospholipase A2 (PLA2), which is expressed as a fusion protein with the Autophagin substrate LC3/ATG8 appended to its N-terminus, as the substrate for the primary enzymatic reaction. The addition of sequences to the N-terminus of PLA2 inhibits the activity of this enzyme. Cleavage by proteases removing the N-terminal extension then restores enzyme activity, constituting the basis for a protease assay. The sub
- Dose response orthogonal assay of uHTS small molecule inhibitors of Striatal-Enriched Phosphatase via a colorimetric intensity assay. Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1R03MH095532-01 Assay Provider: Dr. Lutz Tautz, Sanford-Burnham Medical Research Institute, San Diego CA Disturbance of the dynamic balance between protein tyrosine phosphorylation and dephosphorylation is crucial for the development of many serious conditions, including cancer, diabetes, and autoimmune disorders. This is the first time that tyrosine phosphatase inhibitors are being proposed to improve cognitive function in Alzheimer's disease (AD). STriatal-Enriched Phosphatase (STEP) is a brain-specific protein tyrosine phosphatase that is highly expressed in regions where consolidation of memory occurs and regulates the internalization of NMDARs. Our recent work demonstrates that STEP is elevated in the prefrontal cortex of human AD patients
- Dose responses of compounds that activate the Choline Transporter (CHT) - 10 point CRC Data Source (MLPCN Center Name): Johns Hopkins Ion Channel Center (JHICC) Center Affiliation: Johns Hopkins University, School of Medicine Screening Center PI: Min Li, Ph.D. Assay Provider: Alicia Ruggiero, Ph.D., Vanderbilt University Medical Center Network: Molecular Libraries Probe Production Centers Network (MLPCN) Grant Proposal Number: 1R03DA028852-01 Grant Proposal PI: Alicia Ruggiero, Ph.D., Vanderbilt University Medical Center Assay Implementation: Zhihong Lin Ph. D., Xiaofang Huang M.S., Shunyou Long M.S., Owen McManus Ph.D., and Meng Wu Ph.D. Description: In the brain, the chemical acetylcholine (ACh) exerts powerful modulatory control over arousal, motor and cognitive circuits, and has been found to be deficient in Alzheimer's Disease (AD). The current drugs available to positively impact cognitive deficits in Alzheimer's Disease (AD) and other dementias are the cholinesterase inhibitors. These prevent the breakdown of the neurotransmitter acetylcholine (ACh), an
- Fluorescence Polarization Dose Response Assay for TR3-Based Bcl-B Inhibitors Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA). Network: NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Number: XO1 MH077632-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute This dose response assay is developed and performed for the purpose of confirming hits originally identified in Fluorescence Polarization Screen Assay for Bcl-B Phenotype Converters (AID 1240). Bcl-B is an anti-apoptotic member of the Bcl-2 family that is prominently expressed in plasma and multiple myeloma cells. TR3 (NR4A1; HMR; NP10; GFRP1; NAK1; NUR77; NGFIB) is an orphan member of the steroid/thyroid/retinoid nuclear receptor superfamily that translocates from cellular nuclei to mitochondria upon exposure to various pro-apoptotic stimuli. At mitochondria, TR3 binds to Bcl-B and converts it into a pro-apoptotic protein. A specific 9-amino
- Fluorescence-based biochemical high throughput dose response assay for inhibitors of the Hepatitis C Virus non-structural protein 3 helicase (NS3). Source (MLPCN Center Name): The Scripps Research Institute Molecular Screening Center (SRIMSC) Center Affiliation: The Scripps Research Institute (TSRI) Assay Provider: David Frick, New York Medical College Network: Molecular Libraries Probe Production Centers Network (MLPCN) Grant Proposal Number: 1 R03 MH085690-01 Grant Proposal PI: David Frick, New York Medical College External Assay ID: NS3DNA_INH_FLINT_1536_3XIC50 Name: Fluorescence-based biochemical high throughput dose response assay for inhibitors of the Hepatitis C Virus non-structural protein 3 helicase (NS3). Description: The flavivirus Hepatitis C virus (HCV) is a major cause of liver failure and hepatocellular cancer, with about 170 million people infected worldwide (1). The HCV has a small RNA genome that is directly translated by the infected host cell into a single precursor polyprotein that is processed by enzymatic cleavage into 10 proteins of diverse function. The non-structural proteins include p7, NS2, NS3, NS4
- Fluorescent assay for identification of compounds that inhibit VHR1 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH084230-01A1 Assay Provider: Dr. Lutz Tautz, Sanford-Burnham Medical Research Institute, San Diego CA Protein tyrosine phosphatases (PTPs) play vital roles in numerous cellular processes and are implicated in a growing number of human diseases, ranging from cancer to cardiovascular, immunological, infectious, neurological, and metabolic diseases. The Vaccinia H1-related (VHR) PTP is a dual-specific Erk and Jnk phosphatase, the loss of which causes specific cell cycle arrest in HeLa carcinoma cells, suggesting that VHR inhibition may be a useful approach to halt the growth of cancer cells without detrimental effects on normal cells. Recent studies by collaborators and us suggest that VHR is upregulated in several cervix cancer cell lines, in
- HTS TR-FRET-based dose response confirmatory assay for Siah-1 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH086475-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute, San Diego CA Proteasomal degradation typically requires post-translational modification of target proteins with K48-linked polyubiquitin chains. This process of protein proteolysis plays a key role in normal cellular function. The E3 ubiquitin ligase, Siah-1, facilitates the transfer of ubiquitin to its substrate proteins destined for degradation by way of its RING domain. Siah-1 is a member of a family of highly conserved RING domain proteins, which regulate a variety of cellular functions, including cell cycle arrest, tumor suppression, and apoptosis through the beta-catenin degradation pathway. Siah-1 has also been identified as a p53-inducible gene,
- HTS HePTP Fluorescent Assay using OMFP substrate for In Vitro dose response studies Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Number: XO1 MH077603-01 Assay Provider: Dr. Tomas Mustelin, Sanford-Burnham Medical Research Institute Protein tyrosine phosphatases (PTPs), working with protein tyrosine kinases (PTKs), control the phosphorylation state of many proteins in the signal transduction pathways. HePTP is a tyrosine phosphatase expressed in hematopoietic cells and regulates the MAP kinases Erk and p38. It has been found that HePTP is often dysregualted in the preleukemic disorder myelodysplastic syndrome, as well as in acute myelogeneous leukemia. Small molecule inhibitors of HePTP will be useful as molecular probes for studying the mechanism of signal transduction and MAP kinase regulation, and may have therapeutic potential for the treatment of hematopoietic malignancies. This bioc
- Homogeneous Time-Resolved Fluorescence Resonance Energy Transfer (HTRF) Assay Data Source: Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1 R03 MH085675-01 Assay Provider: Dr Ilya Bezprozvanny, UT Southwestern Medical Center , Dallas, TX Chronic pain (neuropathic pain, inflammatory pain, cancer pain) is a major health problem. Opiate-based drugs, such as morphine and morphine derivatives, are the primary standard of care for the treatment of chronic pain. Unfortunately, patients develop tolerance to opiates due to desensitization of the opiate receptor. Thus, alternative anti-nociceptive ("pain killing") pathways need to be explored for treatment of chronic pain. The N-type voltage-gated Ca2+ channels (CaV2.2s) in dorsal root ganglia neurons is a well validated target for chronic pain (1, 2). We previously demonstrated the interaction between CaV2.2 and the first PDZ domain of molecular
- Image-Based HTS for Selective Antagonists for GPR55 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Proposal Number: 1X01 DA026205-01 Assay Provider: Dr. Mary Abood, California Pacific Medical Center Research Institute The cannabinoid and endocannabinoid system has been implicated in the pathophysiology of drug dependence and addiction disorders. GPR55, an orphan G-Protein Coupled Receptor, has been reported to be a cannabinoid receptor, but its status as such remains unresolved due to conflicting results from pharmacological studies. The goal of this project is to identify small molecule antagonists of GPR55, which may aid in the deorphanization efforts of this receptor and ultimately further the understanding of the role of GPR55 in drug addiction. This high content imaging assay utilizes a cell line permanently expressing a beta-arrestin GFP biosens
- In Vitro Bfl-1 Dose Response Fluorescence Polarization Assay for SAR Study Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Number: XO1 MH077632-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute This Bfl-1 dose response assay is developed and performed for the purpose of SAR study on analogs of hits originally identified in the Bfl-1 fluorescence polarization HTS assay (AID 432). Bfl-1, also known as A1 in mice is an anti-apoptotic and NF-kB-inducible member of the Bcl-2 protein family involved in regulation of apoptosis. Due to difficulties with accomplishing targeted gene ablation in mouse models, the endogenous functions of Bfl-1 are largely unknown. Chemical inhibitors of Bfl-1 can be used as research tools for neutralizing Bfl-1 in human and mouse cells.
- In Vitro Hsp70 Dose Response Fluorescence Polarization Assay for SAR Study Sanford-Burnham Center for Chemical Genomics (SBCCG) Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) NIH Molecular Libraries Screening Centers Network (MLSCN) MLSCN Grant: XO1 MH079863-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute This Hsp70 dose response assay is developed and performed for the purpose of SAR study on analogs of hits originally identified in the fluorescence polarization HTS assay for Hsp70 Inhibitors (AID 583). Over-expression of molecular chaperones occurs commonly in cancers and provides protection from a wide variety of cellular stresses, both endogenous and iatrogenic. Molecular chaperones also play important roles in maintaining the activity of several signal-transducing proteins and transcriptions factors involved in malignant transformation. The human genome contains nine Hsp70-family genes. These chaperones include Hsp70 and Hsc70, which are commonly over-expressed in cancers and which confer resistance
- Luminescent HTS for small molecule inhibitors of MT1-MMP transcription Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Number: XO1 MH78949-01 Assay Provider: Dr. Alex Strongin, Sanford-Burnham Medical Research Institute The sustained presence of matrix metalloproteinases (MMPs) in a tumor environment is a characteristic of many cancer types. The expression of the MT1-MMP mRNA and the MT1-MMP protein closely correlates with increased tumor volume, tumor invasiveness, and the incidence of local and distant metastases. Tumorigenic MT1-MMP is effective in both its active and inactive states. MT1-MMP protects malignant cells against the host immune surveillance thus making tumor cells resistant to the anti-tumor immunity mechanisms. Proteolytically active MT1-MMP is trafficked to centrosomes. Through the proteolysis of centrosomal proteins MT1-MMP promotes mitotic spindle aberrati
- Luminescent assay for HTS discovery of chemical inhibitors of placental alkaline phosphatase confirmation Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Proposal Number: MH077602-01 Assay Provider: Dr. Jose Luis Millan, Sanford-Burnham Medical Research Institute, San Diego, CA. This PLAP dose response assay is developed and performed to confirm hits originally identified in the PLAP Luminescent HTS assay (AID 690) and to study the structure-activity relationship on analogs of the confirmed hits. Compounds are either acquired from commercial sources or synthesized internally. Alkaline phosphatase (EC 3.1.3.1) (APs) catalyze the hydrolysis of phosphomonoesters, releasing phosphate and alcohol. APs are dimeric enzymes found in most organisms. In human, four isozymes of APs have been identified: three isozymes are tissue-specific and the fourth one is tissue-nonspecific. Placental alkaline phosphatase (PLAP) is high
- MOA HePTP Fluorescent secondary assay for identification of redox-state modulating compounds Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Number: XO1 MH077603-01 Assay Provider: Dr. Tomas Mustelin, Sanford-Burnham Medical Research Institute Protein tyrosine phosphatases (PTPs), working with protein tyrosine kinases (PTKs), control the phosphorylation state of many proteins in the signal transduction pathways. HePTP is a tyrosine phosphatase expressed in hematopoietic cells and regulates the MAP kinases Erk and p38. It has been found that HePTP is often dysregualted in the preleukemic disorder myelodysplastic syndrome, as well as in acute myelogeneous leukemia. Small molecule inhibitors of HePTP will be useful as molecular probes for studying the mechanism of signal transduction and MAP kinase regulation, and may have therapeutic potential for the treatment of hematopoietic malignancies. In this a
- MOA VHR1 Fluorescent secondary assay for identification of redox-state modulating compounds Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH084230-01A1 Assay Provider: Dr. Lutz Tautz, Sanford-Burnham Medical Research Institute, San Diego CA Protein tyrosine phosphatases (PTPs) play vital roles in numerous cellular processes and are implicated in a growing number of human diseases, ranging from cancer to cardiovascular, immunological, infectious, neurological, and metabolic diseases. The Vaccinia H1-related (VHR) PTP is a dual-specific Erk and Jnk phosphatase, the loss of which causes specific cell cycle arrest in HeLa carcinoma cells, suggesting that VHR inhibition may be a useful approach to halt the growth of cancer cells without detrimental effects on normal cells. Recent studies by collaborators and us suggest that VHR is upregulated in several cervix cancer cell lines, in
- QFRET-based counterscreen for inhibitors of PFM18AAP: biochemical high throughput dose response assay for inhibitors of the Cathepsin L proteinase (CTSL1). Source (MLPCN Center Name): The Scripps Research Institute Molecular Screening Center Affiliation: The Scripps Research Institute, TSRI Assay Provider: John Dalton and Donald Gardiner, Queensland Institute of Medical Research, Australia Network: Molecular Library Probe Production Centers Network (MLPCN) Grant Proposal Number: 1 R03 MH084103-01 Grant Proposal PI: John Dalton and Donald Gardiner, Queensland Institute of Medical Research, Australia External Assay ID: CTSL1_INH_QFRET_1536_3XIC50 Name: QFRET-based counterscreen for inhibitors of PFM18AAP: biochemical high throughput dose response assay for inhibitors of the Cathepsin L proteinase (CTSL1). Description: Aminopeptidases (APs) are metalloproteases that cleave amino-terminal (N-terminal) amino acids during protein synthesis (1, 2) . These enzymes are characterized in part by their post-translational removal of leucine, aspartate, proline, methionine, etc from proteins and peptides, in order that proteins are properly regulat
- SAR Analysis of Selective Antagonists of GPR55 using an Image-Based Assay - Set 3 Data Source: Sanford-Burnham Center for Chemical Genomics(SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1X01 DA026205-01 Assay Provider: Dr. Mary Abood, (Temple University, formerly at California Pacific Medical Center Research Institute) Addictive behavior stems from abnormal signaling activities in the brain. Thus identification of compounds blocking this modified signaling activity may lead to treatments for addictive behavior. GPR35, a to-date uncharacterized orphan G-Protein Coupled Receptor, is thought to play a role in addiction and has homology to other known receptors of abuse. This high-content imaging assay was used as a counter screen for hits originally identified in a high-content screen for antagonists of the GPR35 receptor "Image-based HTS for Selective Antagonists of GPR35" (AID 2058)) and to study the structure-activity relati
- SAR Analysis of Selective Antagonists of GPR55 using an Image-Based Assay - Set 4 Data Source: Sanford-Burnham Center for Chemical Genomics(SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1X01 DA026205-01 Assay Provider: Dr. Mary Abood, (Temple University, formerly at California Pacific Medical Center Research Institute) The cannabinoid and endocannabinoid system has been implicated in the pathophysiology of drug dependence and addiction disorders. GPR55, an orphan G-Protein Coupled Receptor, has been reported to be a cannabinoid receptor, but its status as such remains unresolved due to conflicting results from pharmacological studies. The goal of the project is to identify small molecule antagonists of GPR55, which may aid in the deorphanization efforts of this receptor and ultimately further the understanding of the role of GPR55 in drug addiction. This high content imaging assay utilizes a cell line permanently expressing
- SAR Analysis of small molecule inhibitors of Sentrin-specific protease 8 (SENP8) using a Luminescent assay - Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network(MLPCN)Grant Proposal Number: 1R21 NS061758-01 fast track Assay Provider: Dr. Guy Salvesen, Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Modification of proteins by SUMO is a dynamic and reversible process. SUMOylation/deSUMOylation cycle regulates SUMOs function. Sentrin-specific proteases (SENPs) are involved in both the maturation of SUMO precursors (endopeptidase cleavage) and deconjugation of the targets (isopeptidase cleavage) [1-3]. There are seven SENPs (1, 2, 3, 5, 6, 7, 8) in humans, and several of these have been characterized as SUMO (or Nedd8) specific enzymes. SENP8 is not a SUMO protease, instead it functions on a small ubiquitin related protein Nedd8. The objective of this project is to generate small molecule inhibitors specific for SENP8
- SAR VHR1 Fluorescent Assay for In Vitro dose response studies Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH084230-01A1 Assay Provider: Dr. Lutz Tautz, Sanford-Burnham Medical Research Institute, San Diego CA Protein tyrosine phosphatases (PTPs) play vital roles in numerous cellular processes and are implicated in a growing number of human diseases, ranging from cancer to cardiovascular, immunological, infectious, neurological, and metabolic diseases. The Vaccinia H1-related (VHR) PTP is a dual-specific Erk and Jnk phosphatase, the loss of which causes specific cell cycle arrest in HeLa carcinoma cells, suggesting that VHR inhibition may be a useful approach to halt the growth of cancer cells without detrimental effects on normal cells. Recent studies by collaborators and us suggest that VHR is upregulated in several cervix cancer cell lines, in
- SAR VHR1 Fluorescent Assay for In Vitro dose response studies Set 3 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH084230-01A1 Assay Provider: Dr. Lutz Tautz, Sanford-Burnham Medical Research Institute, San Diego CA Protein tyrosine phosphatases (PTPs) play vital roles in numerous cellular processes and are implicated in a growing number of human diseases, ranging from cancer to cardiovascular, immunological, infectious, neurological, and metabolic diseases. The Vaccinia H1-related (VHR) PTP is a dual-specific Erk and Jnk phosphatase, the loss of which causes specific cell cycle arrest in HeLa carcinoma cells, suggesting that VHR inhibition may be a useful approach to halt the growth of cancer cells without detrimental effects on normal cells. Recent studies by collaborators and us suggest that VHR is upregulated in several cervix cancer cell lines, in
- SAR VHR1 absorbance Assay for In Vitro dose response studies. Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH084230-01A1 Assay Provider: Dr. Lutz Tautz, Sanford-Burnham Medical Research Institute, San Diego CA Protein tyrosine phosphatases (PTPs) play vital roles in numerous cellular processes and are implicated in a growing number of human diseases, ranging from cancer to cardiovascular, immunological, infectious, neurological, and metabolic diseases. The Vaccinia H1-related (VHR) PTP is a dual-specific Erk and Jnk phosphatase, the loss of which causes specific cell cycle arrest in HeLa carcinoma cells, suggesting that VHR inhibition may be a useful approach to halt the growth of cancer cells without detrimental effects on normal cells. Recent studies by collaborators and us suggest that VHR is upregulated in several cervix cancer cell lines, in
- SAR analysis counterscreen of small molecule antagonists of the CCR6 receptor using an APJ receptor luminescent beta-arrestin assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: R21 NS064746-01A Assay Provider: Dr. Greg Roth, Sanford-Burnham Medical Research Institute This assay was developed and performed as a counter assay for the primary assay originally identified as "uHTS identification of small molecule antagonists of the CCR6 receptor via a luminescent beta-arrestin assay", AID 493098. Compounds are either acquired from commercial sources or synthesized internally. A probe that is selective against APJ (within equipotent range as defined by a >10-fold difference in IC50) is desired. In this description we utilize enzyme-fragment complementation to directly measure GPCR activation. Unlike imaging or other second messenger assays, the DiscoveRx b-Arrestin assay allows for a direct measure of GPCR activation by detect
- SAR analysis of Agonists of the GPR35 Receptor using an Image-Based Assay - Set 3 Data Source: Sanford-Burnham Center for Chemical Genomics Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1X01 DA026205-01 Assay Provider: Dr. Mary Abood, (Temple University, formerly at California Pacific Medical Center Research Institute) Addictive behavior stems from abnormal signaling activities in the brain. Thus identification of compounds blocking this modified signaling activity may lead to treatments for addictive behavior. GPR35, a to-date uncharacterized orphan G-Protein Coupled Receptor, is thought to play a role in addiction and has homology to other known receptors of abuse. This dose response assay is developed and performed to evaluate selectivity of hits originally identified in "Image-Based HTS for Selective Agonists for GPR55" (AID 1961) and to study the structure-activity relationship. Compounds are either acquired from commercial sources or synthesized i
- SAR analysis of Agonists of the GPR35 Receptor using an Image-Based Assay - Set 4 Data Source: Sanford-Burnham Center for Chemical Genomics Source Affiliation: Sanford-Burnham Medical Research Institute Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1X01 DA026205-01 Assay Provider: Dr. Mary Abood, (Temple University, formerly at California Pacific Medical Center Research Institute) Addictive behavior stems from abnormal signaling activities in the brain. Thus identification of compounds blocking this modified signaling activity may lead to treatments for addictive behavior. GPR35, a to-date uncharacterized orphan G-Protein Coupled Receptor, is thought to play a role in addiction and has homology to other known receptors of abuse. This high-content imaging assay is developed and performed to evaluate selectivity of hits originally identified in "Image-based HTS for Selective Antagonists of GPR55" (AID 2013) and to study the structure-activity relationship on analogs of the confirmed hits. Compounds are either acquired fr
- SAR analysis of Antagonists of IAP-family anti-apoptotic proteins - Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network(MLPCN)Grant Proposal Number: MH081277-01 Assay Provider: John C. Reed, Sanford-Burnham Medical Research Institute, San Diego, CA This XIAP dose response assay is developed and performed to confirm hits originally identified in the XIAP HTS binding assay (AID 1018) and to study the structure-activity relationship on analogs of the confirmed hits. Compounds are either acquired from commercial sources or synthesized internally. The assay was performed in the assay providers' laboratory. Apoptosis plays an essential role in many aspects of normal development and physiology, becoming dysregulated in myriad diseases characterized by insufficient or excessive cell death. Caspases are intracellular proteases that are suppressed by Inhibitor of Apoptosis Proteins (IAPs), a famil
- SAR analysis of Antagonists of XIAP-Bir3 domain of IAP-family anti-apoptotic proteins - Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network(MLPCN)Grant Proposal Number: MH081277-01 Assay Provider: John C. Reed, Sanford-Burnham Medical Research Institute San Diego, CA This dose response assay is developed and performed as a counter screen to compounds in the Chemical Antagonists of IAP-family anti-apoptotic proteins confirmation (AID 1449) and to study the structure-activity relationship on analogs of the confirmed hits. Compounds are either acquired from commercial sources or synthesized internally. This assay was performed in the assay providers' laboratory. Apoptosis plays an essential role in many aspects of normal development and physiology, becoming dysregulated in myriad diseases characterized by insufficient or excessive cell death. Caspases are intracellular proteases that are suppressed by Inhibit
- SAR analysis of Antagonists of XIAP-Bir3 domain of IAP-family anti-apoptotic proteins Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Proposal Number: MH081277-01 Assay Provider: John C. Reed, Sanford-Burnham Medical Research Institute, San Diego, CA This dose response assay is developed and performed as a counter screen to compounds in the Chemical Antagonists of IAP-family anti-apoptotic proteins confirmation (AID 1449) and to study the structure-activity relationship on analogs of the confirmed hits. Compounds are either acquired from commercial sources or synthesized internally. This assay was performed in the assay providers' laboratory. Apoptosis plays an essential role in many aspects of normal development and physiology, becoming dysregulated in myriad diseases characterized by insufficient or excessive cell death. Caspases are intracellular proteases that are suppressed by Inhibi
- SAR analysis of Antagonists of the GPR35 Receptor using an Image-Based Assay - Set 4 Data Source: Sanford-Burnham Center for Chemical Genomics Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1X01 DA026205-01 Assay Provider: Dr. Mary Abood, (Temple University, formerly at California Pacific Medical Center Research Institute) Addictive behavior stems from abnormal signaling activities in the brain. Thus identification of compounds blocking this modified signaling activity may lead to treatments for addictive behavior. GPR35, a to-date uncharacterized orphan G-Protein Coupled Receptor, is thought to play a role in addiction and has homology to other known receptors of abuse. This high-content imaging assay is developed and performed to evaluate selectivity of hits originally identified in a high-content screen for agonists of the GPR55 receptor, "Image-Based HTS for Selective Agonists of GPR55"(AID 1961) and to study the structure-activity relatio
- SAR analysis of Antagonists of the GPR35 Receptor using an Image-Based Assay - Set 5 Data Source: Sanford-Burnham Center for Chemical Genomics Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1X01 DA026205-01 Assay Provider: Dr. Mary Abood, (Temple University, formerly at California Pacific Medical Center Research Institute) Addictive behavior stems from abnormal signaling activities in the brain. Thus identification of compounds blocking this modified signaling activity may lead to treatments for addictive behavior. GPR35, a to-date uncharacterized orphan G-Protein Coupled Receptor, is thought to play a role in addiction and has homology to other known receptors of abuse. This high-content imaging assay is developed and performed to confirm activity of hits originally identified in a high-content screen for antagonists of the GPR35 receptor, "Image-Based HTS for Selective Antagonists of GPR35" (AID 2058) and to study the structure-activity
- SAR analysis of Antagonists of the GPR35 Receptor using an Image-Based Assay - Set 6 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1X01 DA026205-01 Assay Provider: Dr. Mary Abood, (Temple University, formerly at California Pacific Medical Center Research Institute) Addictive behavior stems from abnormal signaling activities in the brain. Thus identification of compounds blocking this modified signaling activity may lead to treatments for addictive behavior. GPR35, a to-date uncharacterized orphan G-Protein Coupled Receptor, is thought to play a role in addiction and has homology to other known receptors of abuse. This high-content imaging assay is developed and performed to evaluate selectivity of hits originally identified in a high-content screen for antagonists of the GPR55 receptor, "Image-Based HTS for Selective Antagonists of GPR55" (AID 2013) and to study the struct
- SAR analysis of Antagonists of the GPR35 Receptor using an Image-Based Assay Data Source: Sanford-Burnham Center for Chemical Genomics Source Affiliation: Sanford-Burnham Medical Research Institute Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1X01 DA026205-01 Assay Provider: Dr. Mary Abood, (Temple University, formerly at California Pacific Medical Center Research Institute) Addictive behavior stems from abnormal signaling activities in the brain. Thus identification of compounds blocking this modified signaling activity may lead to treatments for addictive behavior. GPR35, a to-date uncharacterized orphan G-Protein Coupled Receptor, is thought to play a role in addiction and has homology to other known receptors of abuse. This high-content imaging assay is developed and performed to evaluate selectivity of hits originally identified in a high-content screen for agonists of the GPR55 receptor, "Image-Based HTS for Selective Agonists of GPR55"(AID 1961) and to study the structure-activity relationship on analogs of th
- SAR analysis of GM-Tri-DAP induced IL-8 secretion in MCF-7/NOD1 cells - Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Number: 1 R03 MH084844-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute The modulation of immune response activity is one of the major goals in the development of novel therapeutics for auto-immune and inflammatory diseases. The innate system resides at the intersection of the pathways of microbial recognition, inflammation, and cell death, thereby offering various therapeutic targets. In this context, NOD1 and NOD2 are of particular interest, since they recognize distinct structures derived from bacterial peptidoglycans and directly activate NF-kB, a central regulator of immune response, inflammation, and apoptosis. The NOD proteins participate in the signaling events triggered by host recognition of specific motifs (mostly, murope
- SAR analysis of Muramyl dipeptide (MDP) induced IL-8 secretion in MCF-7/NOD2 cells - Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Number: 1 R03 MH084844-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute The modulation of immune response activity is one of the major goals in the development of novel therapeutics for auto-immune and inflammatory diseases. The innate system resides at the intersection of the pathways of microbial recognition, inflammation, and cell death, thereby offering various therapeutic targets. In this context, NOD1 and NOD2 are of particular interest, since they recognize distinct structures derived from bacterial peptidoglycans and directly activate NF-kB, a central regulator of immune response, inflammation, and apoptosis. The NOD proteins participate in the signaling events triggered by host recognition of specific motifs (mostly, murope
- SAR analysis of NF-kB dependent luciferase using DAP as an inducer Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH084844-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute, San Diego CA The modulation of immune response activity is one of the major goals in the development of novel therapeutics for auto-immune and inflammatory diseases. The innate system resides at the intersection of the pathways of microbial recognition, inflammation, and cell death, thereby offering various therapeutic targets. In this context, NOD1 and NOD2 are of particular interest, since they recognize distinct structures derived from bacterial peptidoglycans and directly activate NF-kB, a central regulator of immune response, inflammation, and apoptosis. Mutations in the NOD1 and NOD2 genes are associated with a number of human inflammatory diso
- SAR analysis of NF-kappaB dependent luciferase using DAP as an inducer - Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH084844-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute, San Diego CA The modulation of immune response activity is one of the major goals in the development of novel therapeutics for auto-immune and inflammatory diseases. The innate system resides at the intersection of the pathways of microbial recognition, inflammation, and cell death, thereby offering various therapeutic targets. In this context, NOD1 and NOD2 are of particular interest, since they recognize distinct structures derived from bacterial peptidoglycans and directly activate NF-kB, a central regulator of immune response, inflammation, and apoptosis. Mutations in the NOD1 and NOD2 genes are associated with a number of human inflammatory diso
- SAR analysis of NF-kappaB dependent luciferase using Doxorucibin as an inducer - Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1 R03 MH084844-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute, San Diego CA The modulation of immune response activity is one of the major goals in the development of novel therapeutics for auto-immune and inflammatory diseases. The innate system resides at the intersection of the pathways of microbial recognition, inflammation, and cell death, thereby offering various therapeutic targets. In this context, NOD1 and NOD2 are of particular interest, since they recognize distinct structures derived from bacterial peptidoglycans and directly activate NF-kB, a central regulator of immune response, inflammation, and apoptosis. Mutations in the NOD1 and NOD2 genes are associated with a number of human inflammatory disorders,
- SAR analysis of NF-kappaB dependent luciferase using PMA/Ionomycin as an inducer - 2 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH084844-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute, San Diego CA The modulation of immune response activity is one of the major goals in the development of novel therapeutics for auto-immune and inflammatory diseases. The innate system resides at the intersection of the pathways of microbial recognition, inflammation, and cell death, thereby offering various therapeutic targets. In this context, NOD1 and NOD2 are of particular interest, since they recognize distinct structures derived from bacterial peptidoglycans and directly activate NF-kB, a central regulator of immune response, inflammation, and apoptosis. NF-kB pathway activated by antigen receptors is critical for acquired (as opposed to innate)
- SAR analysis of Tumor necrosis factor alpha (TNF-alpha) induced IL-8 secretion in MCF-7/NOD1 cells - Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Number: 1 R03 MH084844-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute The modulation of immune response activity is one of the major goals in the development of novel therapeutics for auto-immune and inflammatory diseases. The innate system resides at the intersection of the pathways of microbial recognition, inflammation, and cell death, thereby offering various therapeutic targets. In this context, NOD1 and NOD2 are of particular interest, since they recognize distinct structures derived from bacterial peptidoglycans and directly activate NF-kB, a central regulator of immune response, inflammation, and apoptosis. The NOD proteins participate in the signaling events triggered by host recognition of specific motifs (mostly, murope
- SAR analysis of agonists of the Cannabinoid Receptor 1 using an Image-Based Assay - Set 4 Data Source: Sanford-Burnham Center for Chemical Genomics Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1X01 DA026205-01 Assay Provider: Dr. Mary Abood, Temple University, formerly at California Pacific Medical Center Research Institute Drug addiction is a disease that involves the G-protein coupled receptors in the central nervous system resulting in compulsive or abnormal behavior. Recent studies have shown that opioid receptors play a role in regulating other receptors that interact with drug and other substance abuse. The cannabinoid receptors (type 1 and 2) are members of the G-protein coupled receptor family and have been found to be involved in alterations in mood and cognition, as experienced by marijuana users. The specific aim of this assay is to identify small molecule agonists of the human cannabinoid receptor type 1 (CB1). This dose response as
- SAR analysis of agonists of the Cannabinoid Receptor 2 using an Image-Based Assay - Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1X01 DA026205-01 Assay Provider: Dr. Mary Abood, Temple University, formerly at California Pacific Medical Center Research Institute Drug addiction is a disease that involves the G-protein coupled receptors in the central nervous system resulting in compulsive or abnormal behavior. Recent studies have shown that opioid receptors play a role in regulating other receptors that interact with drug and other substance abuse. The cannabinoid receptors (type 1 and 2) are members of the G-protein coupled receptor family and have been found to be involved in alterations in mood and cognition, as experienced by marijuana users. The specific aim of this assay is to identify small molecule agonists of the human cannabinoid receptor type 2 (CB2). This dose response assay
- SAR analysis of agonists of the Cannabinoid Receptor 2 using an Image-Based Assay - Set 3 Data Source: Sanford-Burnham Center for Chemical Genomics Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1X01 DA026205-01 Assay Provider: Dr. Mary Abood, Temple University, formerly at California Pacific Medical Center Research Institute Drug addiction is a disease that involves the G-protein coupled receptors in the central nervous system resulting in compulsive or abnormal behavior. Recent studies have shown that opioid receptors play a role in regulating other receptors that interact with drug and other substance abuse. The cannabinoid receptors (type 1 and 2) are members of the G-protein coupled receptor family and have been found to be involved in alterations in mood and cognition, as experienced by marijuana users. The specific aim of this assay is to identify small molecule agonists of the human cannabinoid receptor type 2 (CB2). This dose respons
- SAR analysis of antagonists of the Cannabinoid Receptor 1 using an Image-Based Assay - Set 3 Data Source: Sanford-Burnham Center for Chemical Genomics Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1X01 DA026205-01 Assay Provider: Dr. Mary Abood, Temple University, formerly at California Pacific Medical Center Research Institute Drug addiction is a disease that involves the G-protein coupled receptors in the central nervous system resulting in compulsive or abnormal behavior. Recent studies have shown that opioid receptors play a role in regulating other receptors that interact with drug and other substance abuse. The cannabinoid receptors (type 1 and 2) are members of the G-protein coupled receptor family and have been found to be involved in alterations in mood and cognition, as experienced by marijuana users. The specific aim of this assay is to identify small molecule antagonists of the human cannabinoid receptor type 1 (CB1). This dose response a
- SAR analysis of antagonists of the Cannabinoid Receptor 1 using an Image-Based Assay - Set 4 Data Source: Sanford-Burnham Center for Chemical Genomics Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1X01 DA026205-01 Assay Provider: Dr. Mary Abood, Temple University, formerly at California Pacific Medical Center Research Institute Drug addiction is a disease that involves the G-protein coupled receptors in the central nervous system resulting in compulsive or abnormal behavior. Recent studies have shown that opioid receptors play a role in regulating other receptors that interact with drug and other substance abuse. The cannabinoid receptors (type 1 and 2) are members of the G-protein coupled receptor family and have been found to be involved in alterations in mood and cognition, as experienced by marijuana users. The specific aim of this assay is to identify small molecule antagonists of the human cannabinoid receptor type 1 (CB1). The dose response ass
- SAR analysis of antagonists of the Cannabinoid Receptor 2 using an Image-Based Assay - Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1X01 DA026205-01 Assay Provider: Dr. Mary Abood, Temple University, formerly at California Pacific Medical Center Research Institute Drug addiction is a disease that involves the G-protein coupled receptors in the central nervous system resulting in compulsive or abnormal behavior. Recent studies have shown that opioid receptors play a role in regulating other receptors that interact with drug and other substance abuse. The cannabinoid receptors (type 1 and 2) are members of the G-protein coupled receptor family and have been found to be involved in alterations in mood and cognition, as experienced by marijuana users. The specific aim of this assay is to identify small molecule antagonists of the human cannabinoid receptor type 2 (CB2). The dose response ass
- SAR analysis of antagonists of the Cannabinoid Receptor 2 using an Image-Based Assay - Set 3 Data Source: Sanford-Burnham Center for Chemical Genomics Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1X01 DA026205-01 Assay Provider: Dr. Mary Abood, Temple University, formerly at California Pacific Medical Center Research Institute Drug addiction is a disease that involves the G-protein coupled receptors in the central nervous system resulting in compulsive or abnormal behavior. Recent studies have shown that opioid receptors play a role in regulating other receptors that interact with drug and other substance abuse. The cannabinoid receptors (type 1 and 2) are members of the G-protein coupled receptor family and have been found to be involved in alterations in mood and cognition, as experienced by marijuana users. The specific aim of this assay is to identify small molecule antagonists of the human cannabinoid receptor type 2 (CB2). The dose respon
- SAR analysis of compounds that inhibit NOD1 - Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH084844-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute, San Diego CA The modulation of immune response activity is one of the major goals in the development of novel therapeutics for auto-immune and inflammatory diseases. The innate system resides at the intersection of the pathways of microbial recognition, inflammation, and cell death, thereby offering various therapeutic targets. In this context, NOD1 and NOD2 are of particular interest, since they recognize distinct structures derived from bacterial peptidoglycans and directly activate NF-kB, a central regulator of immune response, inflammation, and apoptosis. Mutations in the NOD1 and NOD2 genes are associated with a number of human inflammatory diso
- SAR analysis of compounds that inhibit NOD1 - Set 3 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH084844-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute, San Diego CA The modulation of immune response activity is one of the major goals in the development of novel therapeutics for auto-immune and inflammatory diseases. The innate system resides at the intersection of the pathways of microbial recognition, inflammation, and cell death, thereby offering various therapeutic targets. In this context, NOD1 and NOD2 are of particular interest, since they recognize distinct structures derived from bacterial peptidoglycans and directly activate NF-kB, a central regulator of immune response, inflammation, and apoptosis. Mutations in the NOD1 and NOD2 genes are associated with a number of human inflammatory diso
- SAR analysis of compounds that inhibit NOD1 revised Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH084844-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute, San Diego CA The modulation of immune response activity is one of the major goals in the development of novel therapeutics for auto-immune and inflammatory diseases. The innate system resides at the intersection of the pathways of microbial recognition, inflammation, and cell death, thereby offering various therapeutic targets. In this context, NOD1 and NOD2 are of particular interest, since they recognize distinct structures derived from bacterial peptidoglycans and directly activate NF-kB, a central regulator of immune response, inflammation, and apoptosis. Mutations in the NOD1 and NOD2 genes are associated with a number of human inflammatory diso
- SAR analysis of compounds that inhibit NOD2 - Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH084844-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute, San Diego CA The modulation of immune response activity is one of the major goals in the development of novel therapeutics for auto-immune and inflammatory diseases. The innate system resides at the intersection of the pathways of microbial recognition, inflammation, and cell death, thereby offering various therapeutic targets. In this context, NOD1 and NOD2 are of particular interest, since they recognize distinct structures derived from bacterial peptidoglycans and directly activate NF-kB, a central regulator of immune response, inflammation, and apoptosis. Mutations in the NOD1 and NOD2 genes are associated with a number of human inflammatory disor
- SAR analysis of compounds that inhibit NOD2 - Set 3 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH084844-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute, San Diego CA The modulation of immune response activity is one of the major goals in the development of novel therapeutics for auto-immune and inflammatory diseases. The innate system resides at the intersection of the pathways of microbial recognition, inflammation, and cell death, thereby offering various therapeutic targets. In this context, NOD1 and NOD2 are of particular interest, since they recognize distinct structures derived from bacterial peptidoglycans and directly activate NF-kB, a central regulator of immune response, inflammation, and apoptosis. Mutations in the NOD1 and NOD2 genes are associated with a number of human inflammatory disor
- SAR analysis of compounds that inhibit NOD2 revised Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH084844-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute, San Diego CA The modulation of immune response activity is one of the major goals in the development of novel therapeutics for auto-immune and inflammatory diseases. The innate system resides at the intersection of the pathways of microbial recognition, inflammation, and cell death, thereby offering various therapeutic targets. In this context, NOD1 and NOD2 are of particular interest, since they recognize distinct structures derived from bacterial peptidoglycans and directly activate NF-kB, a central regulator of immune response, inflammation, and apoptosis. Mutations in the NOD1 and NOD2 genes are associated with a number of human inflammatory disor
- SAR analysis of compounds that inhibit VHR1, Fluorescent Assay - Set 2 Data Source: Burnham Center for Chemical Genomics (BCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH084230-01A1 Assay Provider: Dr. Lutz Tautz, Source Affiliation: Sanford-Burnham Medical Research Institute (San Diego, CA) Protein tyrosine phosphatases (PTPs) play vital roles in numerous cellular processes and are implicated in a growing number of human diseases, ranging from cancer to cardiovascular, immunological, infectious, neurological, and metabolic diseases. The Vaccinia H1-related (VHR) PTP is a dual-specific Erk and Jnk phosphatase, the loss of which causes specific cell cycle arrest in HeLa carcinoma cells, suggesting that VHR inhibition may be a useful approach to halt the growth of cancer cells without detrimental effects on normal cells. Recent studies by collaborators and us suggest that VHR is upregulated in several cervix cancer ce
- SAR analysis of compounds that potentiate TRAIL-induced apoptosis in MDA-MB-435 cells. Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: X01 MH083230-01 Assay Provider: Dr. Dmitri Rozanov, Sanford-Burnham Medical Research Institute, San Diego CA This assay was developed and performed to confirm hits originally identified in "uHTS for the identification of compounds that potentiate TRAIL-induced apoptosis of cancer cells" (AID 1443) and to study the structure-activity relationship on analogs of the confirmed hits. Compounds are either acquired from commercial sources or synthesized internally. The TRAIL-resistant cell line, MDA-MB-435 is used, because we would like to determine if compounds can potentiate TRAIL-mediated cytotoxicity not only in TRAIL-sensitive PPC-1 carcinoma cells(AIDs 1443 and 1624) but also in TRAIL-resistant cells. Cytotoxic chemotherapy induces apoptosis via a pat
- SAR analysis of inhibitors of TNFa specific NF-kB induction - Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1 R03 MH084844-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute, San Diego CA Multiple cellular stimuli acting through various pathways lead to NF-kB induction. The assay described below uses tumor necrosis factor alpha (TNFa), a canonical NF-kB inducer, and is designed for identification of hits specific to TNFa-modulated pathways. We utilized this assay to assess selectivity of hits emerging from the primary screening of the library in NOD1- and NOD2-specific assays (AIDs 1578 and 1566). The HEK-293-T NF-kB-Luc cell line designed for luminescent detection of NF-kB induction was utilized in this assay. This dose response assay is developed and performed to confirm hits originally identified in "uHTS luminescence assay f
- SAR analysis of inhibitors of TNFa specific NF-kB induction - Set 3 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH084844-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute, San Diego CA Multiple cellular stimuli acting through various pathways lead to NF-kB induction. The assay described below uses tumor necrosis factor alpha (TNFa), a canonical NF-kB inducer, and is designed for identification of hits specific to TNFa-modulated pathways. We utilized this assay to assess selectivity of hits emerging from the primary screening of the library in NOD1- and NOD2-specific assays (AIDs 1578 and 1566). The HEK-293-T NF-kB-Luc cell line designed for luminescent detection of NF-kB induction was utilized in this assay. This dose response assay is developed and performed to confirm hits originally identified in "uHTS luminescence a
- SAR analysis of inhibitors of TNFa specific NF-kB induction revised Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1 R03 MH084844-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute, San Diego CA Multiple cellular stimuli acting through various pathways lead to NF-kB induction. The assay described below uses tumor necrosis factor alpha (TNFa), a canonical NF-kB inducer, and is designed for identification of hits specific to TNFa-modulated pathways. We utilized this assay to assess selectivity of hits emerging from the primary screening of the library in NOD1- and NOD2-specific assays (AIDs 1578 and 1566). The HEK-293-T NF-kB-Luc cell line designed for luminescent detection of NF-kB induction was utilized in this assay. This dose response assay is developed and performed to confirm hits originally identified in "uHTS luminescence assay f
- SAR analysis of small molecule antagonists of the CCR6 receptor: a luminescent beta-arrestin assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: R21 NS064746-01A Assay Provider: Dr. Greg Roth, Sanford-Burnham Medical Research Institute Currently there are no published patents or studies specifically describing small molecule antagonists of the chemokine receptor CCR6. CCL20 (MIP-3 alpha) is the endogenous peptide ligand for the G-protein coupled receptor (GPCR) CCR6. The receptor ligand pair is responsible for the chemoattraction of immature dendritic cells, effector/memory T cells, B cells, and also plays a role at skin and mucosal surfaces. The CCR6 receptor is expressed by B cells, subsets of T cells, and dendritic cells (DC). The link between the CCR6lCCL20 axis and cancer cell metastasis is a recent finding. There are two key studies that describe a relation between CCR6 and colorectal
- SAR analysis of small molecule inhibitors of Mint-PDZ and N-type Ca2+ channel carboxyl-terminal peptide association using HTRF - Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1 R03 MH085675-01 Assay Provider: Dr Ilya Bezprozvanny, UT Southwestern Medical Center , Dallas, TX Chronic pain (neuropathic pain, inflammatory pain, cancer pain) is a major health problem. Opiate-based drugs, such as morphine and morphine derivatives, are the primary standard of care for the treatment of chronic pain. Unfortunately, patients develop tolerance to opiates due to desensitization of the opiate receptor. Thus, alternative anti-nociceptive ("pain killing") pathways need to be explored for treatment of chronic pain. The N-type voltage-gated Ca2+ channels (CaV2.2s) in dorsal root ganglia neurons is a well validated target for chronic pain (1, 2). We previously demonstrated the interaction between CaV2.2 and the first PDZ domain of molecula
- SAR analysis of small molecule inhibitors of Mint-PDZ and N-type Ca2+ channel carboxyl-terminal peptide association using HTRF Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1 R03 MH085675-01 Assay Provider: Dr Ilya Bezprozvanny, UT Southwestern Medical Center , Dallas, TX Chronic pain (neuropathic pain, inflammatory pain, cancer pain) is a major health problem. Opiate-based drugs, such as morphine and morphine derivatives, are the primary standard of care for the treatment of chronic pain. Unfortunately, patients develop tolerance to opiates due to desensitization of the opiate receptor. Thus, alternative anti-nociceptive ("pain killing") pathways need to be explored for treatment of chronic pain. The N-type voltage-gated Ca2+ channels (CaV2.2s) in dorsal root ganglia neurons is a well validated target for chronic pain (1, 2). We previously demonstrated the interaction between CaV2.2 and the first PDZ domain of molecula
- SAR assay for compounds inhibiting TNAP in the absence of phosphate acceptor performed in a luminescent assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Number: MH077602-01 Assay Provider: Dr. Jose Luis Millan, Sanford-Burnham Medical Research Institute, San Diego, CA This TNAP dose response assay is developed and performed for the purpose of SAR study on analogs of hits originally identified in the TNAP luminescent HTS assay (AID 518) Alkaline phosphatase (EC 3.1.3.1) (APs) catalyze the hydrolysis of phosphomonoesters, releasing phosphate and alcohol. APs are dimeric enzymes found in the most organism. In human, four isozymes of APs have been identified. Three isozymes are tissue-specific and the fourth one is tissue-nonsepecifc, named TNAP. TNAP overexpression is associated with excessive calcification observed in different tissues. Therefore, there are therapeutic potentials of in
- SAR assay for compounds that inhibit PHOSPHO1 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH084086-01 Assay Provider: Dr. Jose Luis Milan, Sanford-Burnham Medical Research Institute, San Diego CA Mineralization of cartilage and bone occurs by a series of physicochemical and biochemical processes that together facilitate the deposition of hydroxyapatite (HA) in specific areas of the extracellular matrix (ECM). Experimental evidence has pointed to the presence of HA crystals along collagen fibrils in the ECM and also within the lumen of chondroblast- and osteoblast-derived matrix vesicles (MVs). Dr. Milan's working model is that bone mineralization is first initiated within the lumen of MVs. In a second step, HA crystals grow beyond the confines of the MVs and become exposed to the extracellular milieu where they continue to
- uHTS Homogeneous Terbium Time-Resolved Fluorescence Resonance Energy Transfer (HTRF) Assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH085675-01 Assay Provider: Dr Ilya Bezprozvanny, UT Southwestern Medical Center , Dallas, TX Chronic pain (neuropathic pain, inflammatory pain, cancer pain) is a major health problem. Opiate-based drugs, such as morphine and morphine derivatives, are the primary standard of care for the treatment of chronic pain. Unfortunately, patients develop tolerance to opiates due to desensitization of the opiate receptor. Thus, alternative anti-nociceptive ("pain killing") pathways need to be explored for treatment of chronic pain. The N-type voltage-gated Ca2+ channels (CaV2.2s) in dorsal root ganglia neurons is a well validated target for chronic pain (1, 2). We previously demonstrated the interaction between CaV2.2 and the first PDZ domain of mo
- uHTS absorbance assay for the identification of compounds that inhibit VHR1. Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH084230-01A1 Assay Provider: Dr. Lutz Tautz, Sanford-Burnham Medical Research Institute, San Diego CA Protein tyrosine phosphatases (PTPs) play vital roles in numerous cellular processes and are implicated in a growing number of human diseases, ranging from cancer to cardiovascular, immunological, infectious, neurological, and metabolic diseases. The Vaccinia H1-related (VHR) PTP is a dual-specific Erk and Jnk phosphatase, the loss of which causes specific cell cycle arrest in HeLa carcinoma cells, suggesting that VHR inhibition may be a useful approach to halt the growth of cancer cells without detrimental effects on normal cells. Recent studies by collaborators and us suggest that VHR is upregulated in several cervix cancer cell lines, in
- uHTS luminescence assay for the identification of compounds that inhibit NOD1 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH084844-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute, San Diego CA The modulation of immune response activity is one of the major goals in the development of novel therapeutics for auto-immune and inflammatory diseases. The innate system resides at the intersection of the pathways of microbial recognition, inflammation, and cell death, thereby offering various therapeutic targets. In this context, NOD1 and NOD2 are of particular interest, since they recognize distinct structures derived from bacterial peptidoglycans and directly activate NF-kB, a central regulator of immune response, inflammation, and apoptosis. Mutations in the NOD1 and NOD2 genes are associated with a number of human inflammatory disor
- uHTS luminescence assay for the identification of compounds that inhibit NOD2 in MDP treated cells Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH084844-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute, San Diego, CA The modulation of immune response activity is one of the major goals in the development of novel therapeutics for auto-immune and inflammatory diseases. The innate system resides at the intersection of the pathways of microbial recognition, inflammation, and cell death, thereby offering various therapeutic targets. In this context, NOD1 and NOD2 are of particular interest, since they recognize distinct structures derived from bacterial peptidoglycans and directly activate NF-kB, a central regulator of immune response, inflammation, and apoptosis. Mutations in the NOD1 and NOD2 genes are associated with a number of human inflammatory dis
- Confirmation of compounds inhibiting phosphomannose isomerase (PMI) via a fluorescence intensity assay using a high concentration of mannose 6-phosphate. Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Number: R03 MH082386-01 Assay Provider: Dr. Hudson H. Freeze, Sanford-Burnham Medical Research Institute, San Diego, CA Congenital Disorders of Glycosylation (CDG) are autosomal recessive defects in the synthesis of N-linked oligosaccharide chains. CDG group I (CDG-I) defects are defined as those caused by mutations in genes encoding enzymes used for the synthesis and transfer of lipid linked oligosaccharide (LLO) to newly synthesized proteins in the lumen of the ER. The steps in this pathway and the genes encoding them are very similar from yeast to human. It requires 30-40 single gene products, each dependent on the previous step in the linear sequence to produce and transfer the LLO to protein. Therefore, mutations in any step may cause a type of CDG. There is
- Confirmation of compounds inhibiting phosphomannose isomerase (PMI) via a fluorescence intensity assay. Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Number: R03 MH082386-01 Assay Provider: Dr. Hudson H. Freeze, Sanford-Burnham Medical Research Institute, San Diego, CA Congenital Disorders of Glycosylation (CDG) are autosomal recessive defects in the synthesis of N-linked oligosaccharide chains. CDG group I (CDG-I) defects are defined as those caused by mutations in genes encoding enzymes used for the synthesis and transfer of lipid linked oligosaccharide (LLO) to newly synthesized proteins in the lumen of the ER. The steps in this pathway and the genes encoding them are very similar from yeast to human. It requires 30-40 single gene products, each dependent on the previous step in the linear sequence to produce and transfer the LLO to protein. Therefore, mutations in any step may cause a type of CDG. There is
- Counter Screen for TR3-Based Bcl-B Inhibitors: Fluorescence Polarization Bcl-B/FITC-Bim-BH3 Assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA). Network: NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Number: XO1 MH077632-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute This assay is a counter screen for compounds identified in the Bcl-B/FITC-TR3 fluorescence polarization assay (AID 1240). Bcl-B is an anti-apoptotic member of the Bcl-2 family that is prominently expressed in plasma and multiple myeloma cells. TR3 (also known under NR4A1, HMR, NP10, GFRP1, NAK1, NUR77, and NGFIB) is an orphan member of the steroid/thyroid/retinoid nuclear receptor superfamily that translocates from cellular nuclei to mitochondria upon exposure to various pro-apoptotic stimuli. At mitochondria, TR3 binds to Bcl-B and converts it into a pro-apoptotic protein. A specific 9-amino acid sequence within the full length TR3 is able to
- Dose Response confirmation of uHTS hits for small molecule agonists of the CRF-binding protein and CRF-R2 receptor complex Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Proposal Number: 1 R21 DA029966-01 Assay Providers: Selena Bartlett, Ph.D., Ernest Gallo Clinic and Research Center, University of California, San Francisco and Nick Cosford, Ph.D., Sanford-Burnham Medical Research Institute There is accumulating scientific evidence showing that stressors enhance addictive behaviors and are a common cause of relapse to substance abuse. Corticotrophin releasing factor (CRF) is a 41-aa peptide that has been shown to induce various behavioral changes related to adaptation to stress. The CRF system, including the CRF-binding protein (CRF-BP) and the CRF receptors, CRF-R1 and CRF-R2, are thought to contribute, to the physiological adaptations that result from stress. It has been shown that CRF interaction with CRF-BP may positi
- Dose Response confirmation of uHTS hits from a small molecule agonists of the APJ receptor via a luminescent beta-arrestin assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute(SBMRI, San Diego) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1R21NS059422-01 Assay Provider: Dr. Layton Smith, Sanford-Burnham Medical Research Institute Currently there are no small molecule tools to investigate the biological functions of apelin and its receptor. Apelin is the endogenous peptide ligand for the G-protein coupled receptor (GPCR) APJ (angiotensin II receptor-like 1, AGTRL-1 and APLNR). Until the discovery of apelin, APJ was an orphan GPCR. APJ is coupled to Gai, and has been shown in cell culture to inhibit adenylate cyclase. The APJ gene encodes a receptor that most closely resembles the angiotensin receptor AT1. However, the APJ receptor does not bind angiotensin II. Underscoring the emerging importance of the apelin/APJ system, recent studies have shown that apelin reduces the extent of atherosclerot
- Dose Response confirmation of uHTS hits from a small molecule antagonists of the APJ receptor via a luminescent beta-arrestin assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford- Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1R21NS059422-01 Assay Provider: Dr. Layton Smith, Sanford- Sanford-Burnham Medical Research Institute Currently there are no small molecule tools to investigate the biological functions of apelin and its receptor. Apelin is the endogenous peptide ligand for the G-protein coupled receptor (GPCR) APJ (angiotensin II receptor-like 1, AGTRL-1 and APLNR). Until the discovery of apelin, APJ was an orphan GPCR. APJ is coupled to Gai, and has been shown in cell culture to inhibit adenylate cyclase. The APJ gene encodes a receptor that most closely resembles the angiotensin receptor AT1. However, the APJ receptor does not bind angiotensin II. Underscoring the emerging importance of the apelin/APJ system, recent studies have shown that apelin reduces the
- Dose Response screen for agonists of Angiotensin II Receptor Type 1 to assess selectivity of uHTS small molecule agonists hits of the APJ receptor Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute(SBMRI, San Diego) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1R21NS059422-01 Assay Provider: Dr. Layton Smith, Sanford-Burnham Medical Research Institute Currently there are no small molecule tools to investigate the biological functions of apelin and its receptor. Apelin is the endogenous peptide ligand for the G-protein coupled receptor (GPCR) APJ (angiotensin II receptor-like 1, AGTRL-1 and APLNR). Until the discovery of apelin, APJ was an orphan GPCR. APJ is coupled to Gai, and has been shown in cell culture to inhibit adenylate cyclase. The APJ gene encodes a receptor that most closely resembles the angiotensin receptor AT1. However, the APJ receptor does not bind angiotensin II. Underscoring the emerging importance of the apelin/APJ system, recent studies have shown that apelin reduces the extent of atherosclerot
- Dose Response screen for antagonists of Angiotensin II Receptor Type 1 to assess selectivity of uHTS small molecule antagonists hits of the APJ receptor Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1R21NS059422-01 Assay Provider: Dr. Layton Smith, Sanford-Burnham Medical Research Institute Currently there are no small molecule tools to investigate the biological functions of apelin and its receptor. Apelin is the endogenous peptide ligand for the G-protein coupled receptor (GPCR) APJ (angiotensin II receptor-like 1, AGTRL-1 and APLNR). Until the discovery of apelin, APJ was an orphan GPCR. APJ is coupled to Gai, and has been shown in cell culture to inhibit adenylate cyclase. The APJ gene encodes a receptor that most closely resembles the angiotensin receptor AT1. However, the APJ receptor does not bind angiotensin II. Underscoring the emerging importance of the apelin/APJ system, recent studies have shown that apelin reduces the extent of atherosc
- HTS dose response assay for identification of inhibitors of TNFa-specific NF-kB induction Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1 R03 MH084844-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute, San Diego CA Multiple cellular stimuli acting through various pathways lead to NF-kB induction. The assay described below uses tumor necrosis factor alpha (TNF-a), a canonical NF-kB inducer, and is designed for identification of hits specific to TNF-a-modulated pathways. We utilized this assay to assess selectivity of hits emerging from the primary screening of the library in NOD1- and NOD2-specific assays (AIDs 1578 and 1566). The HEK-293-T NF-kB-Luc cell line designed for luminescent detection of NF-kB induction was utilized in this assay. This assay is the dose response follow-up to "HTS assay for identification of inhibitors of TNF-a-specific NF-kB induc
- HTS fluorescence polarization-based dose response confirmatory screen for the Siah-1 primary assay utilizing an alternative fluorophore, fluorescein-labeled plectin Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH086475-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute, San Diego CA Proteasomal degradation typically requires post-translational modification of target proteins with K48-linked polyubiquitin chains. This process of protein proteolysis plays a key role in normal cellular function. The E3 ubiquitin ligase, Siah-1, facilitates the transfer of ubiquitin to its substrate proteins destined for degradation by way of its RING domain. Siah-1 is a member of a family of highly conserved RING domain proteins, which regulate a variety of cellular functions, including cell cycle arrest, tumor suppression, and apoptosis through the beta-catenin degradation pathway. Siah-1 has also been identified as a p53-inducible gene,
- HTS identification of compounds inhibiting phosphomannose isomerase (PMI) via a fluorescence intensity assay. Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Number: R03 MH082386-01 Assay Provider: Dr. Hudson H. Freeze, Sanford-Burnham Medical Research Institute, San Diego, CA Congenital Disorders of Glycosylation (CDG) are autosomal recessive defects in the synthesis of N-linked oligosaccharide chains. CDG group I (CDG-I) defects are defined as those caused by mutations in genes encoding enzymes used for the synthesis and transfer of lipid linked oligosaccharide (LLO) to newly synthesized proteins in the lumen of the ER. The steps in this pathway and the genes encoding them are very similar from yeast to human. It requires 30-40 single gene products, each dependent on the previous step in the linear sequence to produce and transfer the LLO to protein. Therefore, mutations in any step may cause a type of CDG. There is
- uHTS identification of small molecule antagonists of the binding of Siah-1 and a peptide ligand via a fluorescence polarization assay. Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1 R03 MH086475-01 Assay Provider: Dr. John C. Reed, Sanford-Burnham Medical Research Institute, San Diego CA Proteasomal degradation typically requires post-translational modification of target proteins with K48-linked polyubiquitin chains. This process of protein proteolysis plays a key role in normal cellular function. The E3 ubiquitin ligase, Siah-1, facilitates the transfer of ubiquitin to its substrate proteins destined for degradation by way of its RING domain. Siah-1 is a member of a family of highly conserved RING domain proteins, which regulate a variety of cellular functions, including cell cycle arrest, tumor suppression, and apoptosis through the beta-catenin degradation pathway. Siah-1 has also been identified as a p53-inducible gene, functi
- Counter Screen for Placental Alkaline Phosphatase-based Assays Positives Sanford-Burnham Center for Chemical Genomics (SBCCG) Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Number: MH077609-01 This functional assay was developed for detection of compounds inhibiting placental alkaline phosphatase. These compounds would be observed as false positives of assays employing alkaline phosphatase-based detection. This assay was primarily utilized as counter screen for EphA4 hits identified in the screening assay, AID 689 performed at the Sanford-Burnham Center for Chemical Genomics (SBCCG).
- Receptor Binding Assay In vitro binding assays for the phencyclidine site of NMDA-type glutamate receptors, and for sigma-1 receptors, both of which are known to be bound by DM, were conducted by Sekisui Medical Co., Ltd. of Ibaraki, Japan. In these assays, DM acts as an antagonist at NMDA receptors, which are excitatory in their effects, thereby allowing DM to reduce, calm, and modulate unwanted neuronal activity in neuronal networks that are activated by glutamate agonist activity at NMDA receptors. DM acts as an agonist at sigma-1 receptors, which are inhibitory receptors, thereby allowing DM to reduce unwanted neuronal activity by a second mechanism.
- Enzymatic Assay The aim of this in vitro assay was to measure the inhibition of HCV NS3/4A protease complexes by the compounds of the present invention. This assay provides an indication of how effective compounds of the present invention would be in inhibiting HCV NS3/4A proteolytic activity. The inhibition of full-length hepatitis C NS3 protease enzyme was measured essentially as described in Poliakov, 2002 Prot Expression & Purification 25 363 371. Briefly, the hydrolysis of a depsipeptide substrate, Ac-DED(Edans)EEAbu [COO]ASK(Dabcyl)-NH2 (AnaSpec, San Jose, USA), was measured spectrofluorometrically in the presence of a peptide cofactor, KKGSVVIVGRIVLSGK ( ke Engstrom, Department of Medical Biochemistry and Microbiology, Uppsala University, Sweden). [Landro, 1997 #Biochem 36 9340-9348].
- Inhibition Assay The inhibition of full-length hepatitis C NS3 protease enzyme was measured essentially as described in Poliakov, 2002 Prot Expression & Purification 25 363 371. Briefly, the hydrolysis of a depsipeptide substrate, Ac-DED(Edans)EEAbu[COO]ASK(Dabcyl)-NH2 (AnaSpec, San Jose, USA), was measured spectrofluorometrically in the presence of a peptide cofactor, KKGSVVIVGRIVLSGK (Ake Engstrom, Department of Medical Biochemistry and Microbiology, Uppsala University, Sweden). [Landro, 1997 #Biochem 36 9340-9348]. The enzyme (1 nM) was incubated in 50 mM HEPES, pH 7.5, 10 mM DTT, 40% glycerol, 0.1% n-octyl-D-glucoside, with 25 μM NS4A cofactor and inhibitor at 30 C. for 10 min, whereupon the reaction was initiated by addition of 0.5 μM substrate. Inhibitors were dissolved in DMSO, sonicated for 30 sec. and vortexed.
- Dose response for HTS for Beta-2AR agonists via FAP method from CP1 University of New Mexico Assay Overview: Assay Support: R03 MH093192-01 Project Title: HTS for Non-Canonical Ligands for Beta 2 Adrenergic Receptor Internalization Assay Provider: Jonathan Jarvik, Carnegie Mellon University Screening Center/ PI: UNMCMD/ Larry Sklar Lead Biologist: Yang Wu Chemistry Center/ PI: Vanderbilt Specialty Chemistry Center/Craig Lindsley Chemistry Center Lead: Shaun Stauffer Assay Implementation: Yang Wu, Phillip Tapia, Terry Foutz, Stephanie Chavez, Dominique Perez, Annette Evangelisti, Anna Waller, Cristian Bologa, Mark Carter Assay Background and Significance: G protein-coupled receptors represent the largest family of proteins in the human genome with an estimated number of approximately 800. Because of their central involvement in almost every aspect of human physiology, they also represent the largest target for medical intervention [Lin and Civelli, Annu Med 36 (2004), 204-14]. T
- Dose response for HTS for Beta-2AR agonists via FAP method from Powderset1 University of New Mexico Assay Overview: Assay Support: R03 MH093192-01 Project Title: HTS for Non-Canonical Ligands for Beta 2 Adrenergic Receptor Internalization Assay Provider: Jonathan Jarvik, Carnegie Mellon University Screening Center/ PI: UNMCMD/ Larry Sklar Lead Biologist: Yang Wu Chemistry Center/ PI: Vanderbilt Specialty Chemistry Center/Craig Lindsley Chemistry Center Lead: Shaun Stauffer Assay Implementation: Yang Wu, Phillip Tapia, Terry Foutz, Stephanie Chavez, Dominique Perez, Annette Evangelisti, Anna Waller, Cristian Bologa, Mark Carter Assay Background and Significance: G protein-coupled receptors represent the largest family of proteins in the human genome with an estimated number of approximately 800. Because of their central involvement in almost every aspect of human physiology, they also represent the largest target for medical intervention [Lin and Civelli, Annu Med 36 (2004), 204-14]. T
- Dose response for HTS for Beta-2AR agonists via FAP method from Powderset2 University of New Mexico Assay Overview: Assay Support: R03 MH093192-01 Project Title: HTS for Non-Canonical Ligands for Beta 2 Adrenergic Receptor Internalization Assay Provider: Jonathan Jarvik, Carnegie Mellon University Screening Center/ PI: UNMCMD/ Larry Sklar Lead Biologist: Yang Wu Chemistry Center/ PI: Vanderbilt Specialty Chemistry Center/Craig Lindsley Chemistry Center Lead: Shaun Stauffer Assay Implementation: Yang Wu, Phillip Tapia, Terry Foutz, Stephanie Chavez, Dominique Perez, Annette Evangelisti, Anna Waller, Cristian Bologa, Mark Carter Assay Background and Significance: G protein-coupled receptors represent the largest family of proteins in the human genome with an estimated number of approximately 800. Because of their central involvement in almost every aspect of human physiology, they also represent the largest target for medical intervention [Lin and Civelli, Annu Med 36 (2004), 204-14]. T
- Dose response for HTS for Beta-2AR agonists via FAP method from Powderset3 University of New Mexico Assay Overview: Assay Support: R03 MH093192-01 Project Title: HTS for Non-Canonical Ligands for Beta 2 Adrenergic Receptor Internalization Assay Provider: Jonathan Jarvik, Carnegie Mellon University Screening Center/ PI: UNMCMD/ Larry Sklar Lead Biologist: Yang Wu Chemistry Center/ PI: Vanderbilt Specialty Chemistry Center/Craig Lindsley Chemistry Center Lead: Shaun Stauffer Assay Implementation: Yang Wu, Phillip Tapia, Terry Foutz, Stephanie Chavez, Dominique Perez, Annette Evangelisti, Anna Waller, Cristian Bologa, Mark Carter Assay Background and Significance: G protein-coupled receptors represent the largest family of proteins in the human genome with an estimated number of approximately 800. Because of their central involvement in almost every aspect of human physiology, they also represent the largest target for medical intervention [Lin and Civelli, Annu Med 36 (2004), 204-14]. T
- GAPDH Dose Response Colorimetric Assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Number: None This glyceraldehydes-3-phosphate dehydrogenase (GAPDH; EC 1.2.1.12) dose-response assay is developed and performed at the Sanford-Burnham Center for Chemical Genomics for characterization of the hits of biochemical assays. GAPDH is found in all mammalian tissues, and is considered a "housekeeping enzyme" unaffected by most physiological, hormonal, and metabolic changes. Therefore, compounds that inhibit GAPDH are expected to have pronounced cytotoxic effect and would be unfavorable as chemical probes. In addition, GAPDH has a labile cysteine in its active site that is crucial for the catalysis, resembling several other enzyme groups, such as cysteine-based proteases and phosphatases. As a result, the compounds capable of cysteine oxidation or mod
- HTS Image-Based Screen for Selective Agonists of the KOR Receptor Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak, Duke University, Durham, NC Drug addiction is a disease originating in the central nervous system that produces compulsive behaviors despite the negative consequences that may result. Major addictive drugs of abuse include components of tobacco, opiates, marijuana, ethanol, cocaine, and derivatives of amphetamines. While the addictive behaviors produced by these substances may be generally similar, the drugs act at different receptor sites in the brain. Recent studies have shown that opioid receptors play a role regulating the addictive behaviors of other receptors that interact with illicit and legal substances of abuse. Opioid receptors are composed of multiple subtypes whose contributions to a
- HTS Image-Based Screen for Selective Antagonists of the KOR Receptor Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak , Duke University, Durham NC Drug addiction is a disease originating in the central nervous system that produces compulsive behaviors despite the negative consequences that may result. Major addictive drugs of abuse include components of tobacco, opiates, marijuana, ethanol, cocaine, and derivatives of amphetamines. While the addictive behaviors produced by these substances may be generally similar, the drugs act at different receptor sites in the brain. Recent studies have shown that opioid receptors play a role regulating the addictive behaviors of other receptors that interact with illicit and legal substances of abuse. Opioid receptors are composed of multiple subtypes whose contributions to a
- High Throughput Screening Assay for Hsc70 Inhibitors Sanford-Burnham Center for Chemical Genomics (SBCCG) Sanford-Burnham Medical Research Institute (San Diego, CA) NIH Molecular Libraries Screening Centers Network (MLSCN) Over-expression of molecular chaperones occurs commonly in cancers and provides protection from a wide variety of cellular stresses, both endogenous and iatrogenic. Molecular chaperones also play important roles in maintaining the activity of several signal-transducing proteins and transcriptions factors involved in malignant transformation. The human genome contains nine Hsp70-family genes. These chaperones include Hsp70 and Hsc70, which are commonly over-expressed in cancers and which confer resistance to myriad cellular stresses, including cytotoxic chemotherapy. This work's aim is to identify chemical probes of Hsc70 through a fluorescence polarization assay (FPA) using Fluorescein-labeled ATP. Hsc70 screening was performed at the Sanford-Burnham Center for Chemical Genomics (SBCCG) as part of the Molecular
- High Throughput Screening Assay for Hsp70 Inhibitors Burnham Center for Chemical Genomics (BCCG) Burnham Institute for Medical Research (San Diego, CA) NIH Molecular Libraries Screening Centers Network (MLSCN) MLSCN Grant: XO1 MH079863-01 Over-expression of molecular chaperones occurs commonly in cancers and provides protection from a wide variety of cellular stresses, both endogenous and iatrogenic. Molecular chaperones also play important roles in maintaining the activity of several signal-transducing proteins and transcriptions factors involved in malignant transformation. The human genome contains nine Hsp70-family genes. These chaperones include Hsp70 and Hsc70, which are commonly over-expressed in cancers and which confer resistance to myriad cellular stresses, including cytotoxic chemotherapy. This work's aim is to identify chemical probes of Hsp70 through a fluorescence polarization (FP) assay using Fluorescein-labeled ATP. Additional TR-FRET-based assay was developed and utilized as secondary assay in hit confirmation.
- Inhibition Assay The inhibition of full-length hepatitis C NS3 protease enzyme was measured essentially as described in Poliakov, 2002 Prot Expression & Purification 25 363 371. Briefly, the hydrolysis of a depsipeptide substrate, Ac-DED(Edans)EEAbuψ[COO]ASK(Dabcyl)-NH2 (AnaSpec, San Jos , USA), was measured spectrofluorometrically in the presence of a peptide cofactor, KKGSVVIVGRIVLSGK (Åke Engstr m, Department of Medical Biochemistry and Microbiology, Uppsala University, Sweden) (Landro, 1997 Biochem 36 9340-9348). The enzyme (1 nM) was incubated in 50 mM HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) pH 7.5, 10 mM dithiothreitol, 40% glycerol, 0.1% n-octyl-D-glucoside, with 25 μM NS4A cofactor and inhibitor at 30° C. for 10 min, whereupon the reaction was initiated by addition of 0.5 μM substrate. Inhibitors were dissolved in DMSO, sonicated for 30 sec and vortexed. The solutions were stored at −20° C. between measurements.
- Luminescent assay for HTS discovery of chemical activators of placental alkaline phosphatase Sanford-Burnham Center for Chemical Genomics (SBCCG) Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) NIH Molecular Libraries Screening Centers Network (MLSCN) Alkaline phosphatase (EC 3.1.3.1) (APs) catalyze the hydrolysis of phosphomonoesters, releasing phosphate and alcohol. APs are dimeric enzymes found in most organisms. In human, four isozymes of APs have been identified: three isozymes are tissue-specific and the fourth one is tissue-nonspecific. Placental alkaline phosphatase (PLAP) is highly expressed in primate placental tissue. Its biological function is unknown. Identification of PLAP-specific inhibitors should provide necessary tools for characterization of its biological role. PLAP screening was developed and performed at the Sanford-Burnham Center for Chemical Genomics (SBCCG) as part of the Molecular Library Screening Center Network (MLSCN). This assay represents a selectivity screening for tissue nonspecific alkaline phosphatase (TNAP) screened at BCC
- Luminescent assay for HTS discovery of chemical inhibitors of placental alkaline phosphatase Sanford-Burnham Center for Chemical Genomics (SBCCG) Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) NIH Molecular Libraries Screening Centers Network (MLSCN) Alkaline phosphatase (EC 3.1.3.1) (APs) catalyze the hydrolysis of phosphomonoesters, releasing phosphate and alcohol. APs are dimeric enzymes found in most organisms. In human, four isozymes of APs have been identified: three isozymes are tissue-specific and the fourth one is tissue-nonspecific. Placental alkaline phosphatase (PLAP) is highly expressed in primate placental tissue. Its biological function is unknown. Identification of PLAP-specific inhibitors should provide necessary tools for characterization of its biological role. PLAP screening was developed and performed at the Sanford-Burnham Center for Chemical Genomics (SBCCG) as part of the Molecular Library Screening Center Network (MLSCN). This assay represents a selectivity screening for tissue nonspecific alkaline phosphatase (TNAP) screened at B
- SAR analysis for the identification of translation initiation inhibitors (PABP) Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1R03MH084835-01 Assay Provider: Jerry Pelletier, Ph.D, McGill University, Montreal, Canada Translation is an essential cellular process whose deregulation is associated with alterations in cell growth, cell cycle progression, and cell death responses. The initiation phase of translation is a key target for regulation when cells are exposed to various environmental cues (e.g. insulin, amino acid starvation, mitogenic stimulation, hypoxia, etc). As well, translation initiation control is usurped upon viral infection and is deregulated in many human cancers. Over-expression of certain translation factors can lead to malignant transformation and many of the components of the translational apparatus are over-expressed in human cancers. Several tumor sup
- SAR analysis for the identification of translation initiation inhibitors (eIF4H) Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBIMR, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1R03MH084835-01 Assay Provider: Jerry Pelletier, Ph.D, McGill University, Montreal, Canada Translation is an essential cellular process whose deregulation is associated with alterations in cell growth, cell cycle progression, and cell death responses. The initiation phase of translation is a key target for regulation when cells are exposed to various environmental cues (e.g. insulin, amino acid starvation, mitogenic stimulation, hypoxia, etc). As well, translation initiation control is usurped upon viral infection and is deregulated in many human cancers. Over-expression of certain translation factors can lead to malignant transformation and many of the components of the translational apparatus are over-expressed in human cancers. Several tumor su
- TNAP luminescent HTS assay Sanford-Burnham Center for Chemical Genomics (SBCCG) Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Number: MH077602-01 Alkaline phosphatase (EC 3.1.3.1) (APs) catalyze the hydrolysis of phosphomonoesters, releasing phosphate and alcohol. APs are dimeric enzymes found in the most organism. In human, four isozymes of APs have been identified. Three isozymes are tissue-specific and the fourth one is tissue-nonsepecifc, named TNAP. TNAP deficiency is associated with defective bone mineralization in the form of rickets and osteomalacia. Therefore, there are therapeutic potentials of inhibiting TNAP activity. The goal of this HTS is to identify novel and specific inhibitors of TNAP. TNAP screening was developed and performed at the Sanford-Burnham Center for Chemical Genomics (SBCCG) as part of the Molecular Library Screening Center Network (MLSCN). XO1 submission, MH077602-01, Pharmacological inhibitors o
- uHTS fluorescence polarization assay for the identification of translation initiation inhibitors (PABP) Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1R03MH084835-01 Assay Provider: Jerry Pelletier, Ph.D, McGill University, Montreal, Canada Translation is an essential cellular process whose deregulation is associated with alterations in cell growth, cell cycle progression, and cell death responses. The initiation phase of translation is a key target for regulation when cells are exposed to various environmental cues (e.g. insulin, amino acid starvation, mitogenic stimulation, hypoxia, etc). As well, translation initiation control is usurped upon viral infection and is deregulated in many human cancers. Over-expression of certain translation factors can lead to malignant transformation and many of the components of the translational apparatus are over-expressed in human cancers. Several tumor suppresso
- uHTS fluorescence polarization assay for the identification of translation initiation inhibitors (eIF4H) Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1R03MH084835-01 Assay Provider: Jerry Pelletier, Ph.D, McGill University, Montreal, Canada Translation is an essential cellular process whose deregulation is associated with alterations in cell growth, cell cycle progression, and cell death responses. The initiation phase of translation is a key target for regulation when cells are exposed to various environmental cues (e.g. insulin, amino acid starvation, mitogenic stimulation, hypoxia, etc). As well, translation initiation control is usurped upon viral infection and is deregulated in many human cancers. Over-expression of certain translation factors can lead to malignant transformation and many of the components of the translational apparatus are over-expressed in human cancers. Several tumor suppresso
- AlphaScreen confirmatory assay for validation of inhibitors of SUMOylation Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH084862-01 Assay Provider: Dr. Yuan Chen, Beckman Research Institute, City Of Hope, CA Protein modification by the SUMO (Small Ubiquitin-like MOdifier) family of proteins is an important post-translational modification that plays an essential role in many functions including gene transcription, cell cycle progression, DNA repair, viral infection, and the development of neurodegenerative diseases (1, 2). Recent proteomic studies have found that approximately 10% of the proteins encoded by the yeast genome are substrates for SUMO modification (3-5). The mechanism of how SUMOylation is involved in these cellular functions remains largely unclear. The inhibitors of SUMOylation would be useful to probe the roles of SUMOylation in cellular regulat
- Beta2-AR Binding Assay HEK 293 cells stability transfected with cDNA encoding human beta2-AR (provided by Dr. Brian Kobilka, Stanford Medical Center, Palo Alto, Calif.) were grown in Dulbecco's Modified Eagle Medium (DMEM) containing 10% fetal bovine serum (FBS), 0.05% penicillin-streptomycin, and 400 g/ml G418 as previously described (Pauwels et al., Biochem. Pharmacol. 42: 1683-1689, 1991). The cells were scraped from the 150x25 mm plates and centrifuged at 500xg for 5 minutes. The pellet was homogenized in 50 mM Tris-HCl, pH 7.7, with a Polytron, centrifuged at 27,000xg, and resuspended in the same buffer. The latter process was repeated, and the pellet was resuspended in 25 mM Tris-HCl containing 120 mM NaCl, 5.4 mM KCl, 1.8 mM CaCl2, 0.8 mM MgCl2, and 5 mM glucose, pH 7.4. The binding assays contained 0.3 nM [3H]CGP-12177 in a volume of 1.0 ml. Nonspecific binding was determined by 1 uM propranolol.
- Dose Response concentration confirmation of uHTS hits from a small molecule activators of mouse intestinal alkaline phosphatase via a luminescent assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Proposal Number: X01-MH077602-01 Alkaline phosphatase (EC 3.1.3.1) (APs) catalyze the hydrolysis of phosphomonoesters, releasing inorganic phosphate and alcohol. APs are dimeric enzymes found in most organisms. In human, four isozymes of APs have been identified. One isozyme is tissue-nonspecific (designated TNAP) and three other isozymes are tissue-specific and named according to the tissue of their predominant expression: intestinal (IAP), placental (PLAP) and germ cell (GCAP) alkaline phosphatases. IAP expression is largely restricted to the gut, especially to the epithelial cells (enterocytes) of the small intestinal mucosa. The exact biological function of IAP is unknown. The goal of this HTS is to confirm hits in "uHTS Luminescent assay for identificatio
- Dose Response confirmation of Inhibitors of Mdm2/MdmX interaction in luminescent format Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: R03 MH089489-01 Assay Provider: Dr. Geoffrey M. Wahl, Salk Institute for Biological Studies, San Diego, CA A wild type but attenuated p53 is retained in approximately 50% of human tumors, and reactivation of p53 in such tumors is an attractive chemotherapeutic strategy. p53 activity is restricted in vivo by mdm2 and mdmx, and knockout of either of these proteins is embryonic lethal in a p53-dependent manner (1, 2). Both proteins bind to p53 via a hydrophobic N-terminal pocket and block p53-dependent transcription of genes required for tumor suppression. Efforts to reactivate p53 with small molecules have focused on inhibition of the mdm2/p53 interaction, which leads to increased p53 levels and activity. However, recent reports indicate that targetin
- Dose Response confirmation of activators of hexokinase domain containing I (HKDC1) Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Proposal Number: R21 NS061703-01S1 Assay Provider: Dr. Jeff Johnson, Metabolex, Inc., Hayward, CA. The gene HKDC1 (HexoKinase Domain Containing I) was found to encode a fifth mammalian hexokinase. Since hexokinases (HKs) are the first step in glucose metabolism, they play a major role in regulating the metabolic fate of glucose in the tissues they are expressed. Understanding both the proper and pathological metabolism of glucose is obviously of critical importance to deciphering the dysregulation of glucose that leads to Type 2 diabetes, a disorder of metabolism that morbidly afflicts 35 million Americans and perhaps close to 200 million worldwide. It is our expectation that HTS to find small molecule activators for HKDC1 will begin the process of identif
- Dose Response confirmation of activators of hexokinase domain containing I (HKDC1) in the hexokinase 1 selectivity assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Proposal Number: R21 NS061703-01S1 Assay Provider: Dr. Jeff Johnson, Metabolex, Inc., Hayward, CA. Since hexokinases (HKs) are the first step in glucose metabolism, they play a major role in regulating the metabolic fate of glucose in the tissues they are expressed. Understanding both the proper and pathological metabolism of glucose is obviously of critical importance to deciphering the dysregulation of glucose that leads to Type 2 diabetes, a disorder of metabolism that morbidly afflicts 35 million Americans and perhaps close to 200 million worldwide. One member of HK enzyme family, Hexokinase IV or glucokinase, has been the subject of intense pharmaceutical development of small molecule activators for treatment of Type 2 diabetes. The other members of
- Dose Response confirmation of inhibitors of hexokinase domain containing I (HKDC1) Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Proposal Number: R21 NS061703-01S1 Assay Provider: Dr. Jeff Johnson, Metabolex, Inc., Hayward, CA. The gene HKDC1 (HexoKinase Domain Containing I) was found to encode a fifth mammalian hexokinase. Since hexokinases are the first step in glucose metabolism, they play a major role in regulating the metabolic fate of glucose in the tissues they are expressed. Understanding both the proper and pathological metabolism of glucose is obviously of critical importance to deciphering the dysregulation of glucose metabolism that leads to Type 2 diabetes, a disorder of glucose metabolism that morbidly afflicts 35 million Americans and perhaps close to 200 million worldwide. It is our expectation that HTS to find small molecule inhibitors for HKDC1 will begin the proce
- Dose Response confirmation of inhibitors of hexokinase domain containing I (HKDC1) in the hexokinase 1 selectivity assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Proposal Number: R21 NS061703-01S1 Assay Provider: Dr. Jeff Johnson, Metabolex, Inc., Hayward, CA. Since hexokinases (HKs) are the first step in glucose metabolism, they play a major role in regulating the metabolic fate of glucose in the tissues they are expressed. Understanding both the proper and pathological metabolism of glucose is obviously of critical importance to deciphering the dysregulation of glucose that leads to Type 2 diabetes, a disorder of metabolism that morbidly afflicts 35 million Americans and perhaps close to 200 million worldwide. The gene HKDC1 (HexoKinase Domain Containing I) was found to encode a fifth mammalian hexokinase. Its expression pattern is unique among hexokinases, providing some initial clues to its function in biolog
- Dose Response confirmation of uHTS Activators of the Apaf-1 Pathway in Fluorescent format Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBIMR, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: R01 CA136513 Assay Provider: Dr. Xuejun Jiang, Sloan-Kettering Institute for Cancer Research, New York, NY Apoptosis is a major form of programmed cell death that multicellular organisms utilize to maintain tissue homeostasis and to eliminate unwanted or damaged cells. It plays a critical role in development, immune responses and many other physiological events. In mammals, the mitochondrial cytochrome c-mediated apoptotic pathway is initiated by cytochrome c release from mitochondria. Deregulation of the cytochrome c apoptotic pathway can lead to diseases such as cancer, immune disorders, and neurodegenerative diseases. The overall goal of this project is to identify Apaf-1 activators by high throughput screening. Apaf-1 is the essential mediato
- Dose Response confirmation of uHTS activators of Human Intestinal Alkaline Phosphatase using Mouse Intestinal Alkaline Phosphatase Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Proposal Number: X01-MH077602-01 Alkaline phosphatase (EC 3.1.3.1) (APs) catalyze the hydrolysis of phosphomonoesters, releasing inorganic phosphate and alcohol. APs are dimeric enzymes found in most organisms. In human, four isozymes of APs have been identified. One isozyme is tissue-nonspecific (designated TNAP) and three other isozymes are tissue-specific and named according to the tissue of their predominant expression: intestinal (IAP), placental (PLAP) and germ cell (GCAP) alkaline phosphatases. IAP expression is largely restricted to the gut, especially to the epithelial cells (enterocytes) of the small intestinal mucosa. The exact biological function of IAP is unknown. The goal of this MLPCN probe project is to identify novel and specific activators of
- Dose Response confirmation of uHTS activators of Mouse Intestinal Alkaline Phosphatase using Placental Alkaline Phosphatase Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Proposal Number: X01-MH077602-01 Alkaline phosphatase (EC 3.1.3.1) (APs) catalyze the hydrolysis of phosphomonoesters, releasing inorganic phosphate and alcohol. APs are dimeric enzymes found in most organisms. In human, four isozymes of APs have been identified. One isozyme is tissue-nonspecific (designated TNAP) and three other isozymes are tissue-specific and named according to the tissue of their predominant expression: intestinal (IAP), placental (PLAP) and germ cell (GCAP) alkaline phosphatases. IAP expression is largely restricted to the gut, especially to the epithelial cells (enterocytes) of the small intestinal mucosa. The exact biological function of IAP is unknown. The goal of this HTS is to confirm hits in "uHTS Luminescent assay for identificatio
- Dose Response confirmation of uHTS activators of Mouse Intestinal Alkaline Phosphatase using Tissue Nonspecific Alkaline Phosphatase. Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Proposal Number: X01-MH077602-01 Alkaline phosphatase (EC 3.1.3.1) (APs) catalyze the hydrolysis of phosphomonoesters, releasing inorganic phosphate and alcohol. APs are dimeric enzymes found in most organisms. In human, four isozymes of APs have been identified. One isozyme is tissue-nonspecific (designated TNAP) and three other isozymes are tissue-specific and named according to the tissue of their predominant expression: intestinal (IAP), placental (PLAP) and germ cell (GCAP) alkaline phosphatases. IAP expression is largely restricted to the gut, especially to the epithelial cells (enterocytes) of the small intestinal mucosa. The exact biological function of IAP is unknown. The goal of this HTS is to confirm hits in "uHTS Luminescent assay for identificatio
- Dose Response confirmation of uHTS hits for Scp-1 phosphatase using a colorimetric assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 DA030556-01A1 Assay Provider: Dr. Yan Jessie Zhang, University of Texas, Austin, TX Human Scp phosphatases are regulators involved in neuronal gene silencing. This family of phosphatases exhibits specificity for phosphoryl-Ser5 in the heptad repeats of the C-terminal domain of RNA polymerase II (RNA Pol II) and thus inhibit initiation of transcription [1, 2]. Scps strongly associate with the REST/NRSF neuronal silencing complex that functions to inhibit transcription of neuronal genes in neuronal stem cells and in nonneuronal tissues [1]. Inhibition of Scps by dominant negative forms of the enzyme or by premature expression of the nervous system-specific microRNA miR-124, results in differentiation of neuronal stem cells into mature neurons [
- Dose Response confirmation of uHTS hits from a small molecule inhibitors of LYP via a fluorescence intensity assay - Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1R21NS056945-01 Assay Provider: Dr. Nunzio Bottini, La Jolla Institute for Allergy and Immunology, La Jolla, CA LYP, is a lymphocyte specific protein tyrosine phosphatase that plays a critical regulatory role in T cell receptor signaling. The PTPN22 gene encodes this phosphatase. A single-nucleotide polymorphism in PTPN22 is associated with a number of autoimmune disorders, including type 1 diabetes, rheumatoid arthritis, juvenile rheumatoid arthritis, systemic lupus erythematosus and Grave's disease. The autoimmunity-predisposing allele is a gain-of-function mutant suggesting that a specific small-molecule inhibitor could eliminate its effect. This biochemical assay employs a fluorescent readout based on the enzyme's ability to liberate
- Dose Response confirmation of uHTS hits from a small molecule inhibitors of LYP via a fluorescence intensity assay using pCAP substrate Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1R21NS056945-01 Assay Provider: Dr. Nunzio Bottini, La Jolla Institute for Allergy and Immunology, La Jolla, CA LYP, is a lymphocyte specific protein tyrosine phosphatase that plays a critical regulatory role in T cell receptor signaling. The PTPN22 gene encodes this phosphatase. A single-nucleotide polymorphism in PTPN22 is associated with a number of autoimmune disorders, including type 1 diabetes, rheumatoid arthritis, juvenile rheumatoid arthritis, systemic lupus erythematosus and Grave's disease. The autoimmunity-predisposing allele is a gain-of-function mutant suggesting that a specific small-molecule inhibitor could eliminate its effect. Finding specific inhibitors of protein phosphatases has proven extremely difficult. The goal of th
- Dose Response confirmation of uHTS small molecule inhibitors of Plasmodium falciparum Glucose-6-phosphate dehydrogenase via a fluorescence intensity assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1R21AI082434-01 Assay Provider: Lars Bode, Ph.D., University of California San Diego, San Diego, CA Tropical malaria caused by the protozoan parasite Plasmodium falciparum is responsible for up to three million deaths annually. Due to increasing regional distribution and resistances against the clinically used antimalarials, novel antimalarial drugs - which have new mechanisms of action and are suitable for combination therapies - are urgently required. Plasmodium falciparum Glucose-6-phosphate dehydrogenase (PfGluPho) is a potential novel target for antimalarial drug design. Glucose-6-Phosphate Dehydrogenase (G6PD) reaction is the first and rate-limiting step in the pentose phosphate pathway (PPP), catalyzed by a bifunctional enzyme Plasmodium fal
- Dose Response confirmation of uHTS small molecule inhibitors of tim10-1 yeast via a luminescent assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 DA027714-01A1 Assay Provider: Dr. Carla Koehler, University of California, Los Angeles, CA (UCLA) Defects in mitochondrial assembly impact a wide range of diseases from degenerative muscle and neural diseases to cancer (Wallace, 2005). The mitochondrion is not only important for the production of energy but plays an important role in other aspects such as intermediary metabolism and signaling. The mitochondrion contains an inner membrane and outer membrane that separate the matrix from the intermembrane space. Proteins destined for the mitochondrion are imported via the Translocase of the Outer Membrane (TOM) and the Translocases of the Inner Membrane (TIM23 for proteins destined for the matrix and TIM22 for proteins destined for the inner me
- Dose Response confirmation of uHTS small molecule inhibitors of tim10-1: a luminescent tim23-1 yeast counterscreen. Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 DA027714-01A1 Assay Provider: Dr. Carla Koehler, University of California, Los Angeles, CA (UCLA) Defects in mitochondrial assembly impact a wide range of diseases from degenerative muscle and neural diseases to cancer (Wallace, 2005). The mitochondrion is not only important for the production of energy but plays an important role in other aspects such as intermediary metabolism and signaling. The mitochondrion contains an inner membrane and outer membrane that separate the matrix from the intermembrane space. Proteins destined for the mitochondrion are imported via the Translocase of the Outer Membrane (TOM) and the Translocases of the Inner Membrane (TIM23 for proteins destined for the matrix and TIM22 for proteins destined for the inner me
- Dose Response confirmation of uHTS small molecule inhibitors of tim23-1: a luminescent TIM10 yeast counterscreen. Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 DA027714-01A1 Assay Provider: Dr. Carla Koehler, University of California, Los Angeles, CA (UCLA) Defects in mitochondrial assembly impact a wide range of diseases from degenerative muscle and neural diseases to cancer (Wallace, 2005). The mitochondrion is not only important for the production of energy but plays an important role in other aspects such as intermediary metabolism and signaling. The mitochondrion contains an inner membrane and outer membrane that separate the matrix from the intermembrane space. Proteins destined for the mitochondrion are imported via the Translocase of the Outer Membrane (TOM) and the Translocases of the Inner Membrane (TIM23 for proteins destined for the matrix and TIM22 for proteins destined for the inner me
- Dose Response confirmation of uHTS small molecule inhibitors of tim23-1: a luminescent tim10-1 yeast counterscreen. Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 DA027714-01A1 Assay Provider: Dr. Carla Koehler, University of California, Los Angeles, CA (UCLA) Defects in mitochondrial assembly impact a wide range of diseases from degenerative muscle and neural diseases to cancer (Wallace, 2005). The mitochondrion is not only important for the production of energy but plays an important role in other aspects such as intermediary metabolism and signaling. The mitochondrion contains an inner membrane and outer membrane that separate the matrix from the intermembrane space. Proteins destined for the mitochondrion are imported via the Translocase of the Outer Membrane (TOM) and the Translocases of the Inner Membrane (TIM23 for proteins destined for the matrix and TIM22 for proteins destined for the inner me
- Dose Response orthogonal assay utilizing the direct end-point detection of NADPH for uHTS small molecule inhibitors of Plasmodium falciparum Glucose-6-phosphate dehydrogenase Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1R21AI082434-01 Assay Provider: Lars Bode, Ph.D., University of California San Diego, San Diego, CA Tropical malaria caused by the protozoan parasite Plasmodium falciparum is responsible for up to three million deaths annually. Due to increasing regional distribution and resistances against the clinically used antimalarials, novel antimalarial drugs - which have new mechanisms of action and are suitable for combination therapies - are urgently required. Plasmodium falciparum Glucose-6-phosphate dehydrogenase (PfGluPho) is a potential novel target for antimalarial drug design. Glucose-6-Phosphate Dehydrogenase (G6PD) reaction is the first and rate-limiting step in the pentose phosphate pathway (PPP), catalyzed by a bifunctional enzyme Plasmodium fal
- Dose Response orthogonal kinetic assay utilizing the direct detection of NADPH for uHTS small molecule inhibitors of Plasmodium falciparum Glucose-6-phosphate dehydrogenase Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1R21AI082434-01 Assay Provider: Lars Bode, Ph.D., University of California San Diego, San Diego, CA Tropical malaria caused by the protozoan parasite Plasmodium falciparum is responsible for up to three million deaths annually. Due to increasing regional distribution and resistances against the clinically used antimalarials, novel antimalarial drugs - which have new mechanisms of action and are suitable for combination therapies - are urgently required. Plasmodium falciparum Glucose-6-phosphate dehydrogenase (PfGluPho) is a potential novel target for antimalarial drug design. Glucose-6-Phosphate Dehydrogenase (G6PD) reaction is the first and rate-limiting step in the pentose phosphate pathway (PPP), catalyzed by a bifunctional enzyme Plasmodium fal
- Dose Response validation of Activators of Apaf-1 using a Fluorescent Interference Counterscreen assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBIMR, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: R01 CA136513 Assay Provider: Dr. Xuejun Jiang, Sloan-Kettering Institute for Cancer Research, New York, NY Apoptosis is a major form of programmed cell death that multicellular organisms utilize to maintain tissue homeostasis and to eliminate unwanted or damaged cells. It plays a critical role in development, immune responses and many other physiological events. In mammals, the mitochondrial cytochrome c-mediated apoptotic pathway is initiated by cytochrome c release from mitochondria. Deregulation of the cytochrome c apoptotic pathway can lead to diseases such as cancer, immune disorders, and neurodegenerative diseases. The overall goal of this project is to identify Apaf-1 activators by high throughput screening. Apaf-1 is the essential mediato
- Dose Response validation of Inhibitors of Apaf-1 using a Fluorescent Interference Counterscreen assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBIMR, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: R01 CA136513 Assay Provider: Dr. Xuejun Jiang, Sloan-Kettering Institute for Cancer Research, New York, NY Apoptosis is a major form of programmed cell death that multicellular organisms utilize to maintain tissue homeostasis and to eliminate unwanted or damaged cells. It plays a critical role in development, immune responses and many other physiological events. In mammals, the mitochondrial cytochrome c-mediated apoptotic pathway is initiated by cytochrome c release from mitochondria. Deregulation of the cytochrome c apoptotic pathway can lead to diseases such as cancer, immune disorders, and neurodegenerative diseases. The overall goal of this project is to identify Apaf-1 inhibitors by high throughput screening. Apaf-1 is the essential mediato
- Dose Response validation of uHTS RPN11 inhibitor hits using a Thrombin Fluorescence Polarization assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford Burnham Medical Research Institute (SBMRI, La Jolla, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH094180-01 Assay Provider: Dr. Raymond Deshaies, California Institute of Technology, Pasadena, CA Protein modification by the attachment of ubiquitin to cellular proteins is a key mechanism in regulating many cellular and physiological processes. Ubiquitin is covalently attached via an enzymatic cascade to target proteins through an isopeptide bond between the C-terminus of ubiquitin and a lysine residue of the acceptor substrate [1]. Assembly of a chain of >=4 ubiquitins linked together via Lys48 of ubiquitin marks cellular proteins for degradation by the 26S proteasome [2-3]. The 26S proteasome is a 2.5 megadalton macromolecular protein complex that comprises two distinct subparticles: the 19S cap regulatory particle (RP) and the 20S core
- Dose response confirmation of DNMT1 inhibitors in a Fluorescent Molecular Beacon assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Proposal Number: 1 R03 DA031091-01A1 Assay Provider: Dr. James Stivers, Johns Hopkins University, Baltimore, MD Hypermethylation of CpG dinucleotides in promoter regions of tumor suppressor genes by DNA 5-C-MTases is an important hallmark of human cancers [1-3]. Such reversible epigenetic silencing is a key pathway resulting in loss-of-function phenotypes that promote the growth of many cancers. Examples of genes that have undergone hypermethylation-induced silencing include the genes encoding the cell cycle regulator proteins pIS and p16, the pro-apoptotic effector gene Apaf-1, the mismatch DNA repair gene MLH1, and GSTP1 that codes for the phase 2 enzyme glutathione S-transferase [1, 2, 4-9]. Thus, inhibitors of MTase enzymes have the demonstrated potent
- Dose response confirmation of uHTS RPN11 inhibitor hits in a Fluorescence Polarization assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford Burnham Medical Research Institute (SBMRI, La Jolla, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH094180-01 Assay Provider: Dr. Raymond Deshaies, California Institute of Technology, Pasadena, CA Protein modification by the attachment of ubiquitin to cellular proteins is a key mechanism in regulating many cellular and physiological processes. Ubiquitin is covalently attached via an enzymatic cascade to target proteins through an isopeptide bond between the C-terminus of ubiquitin and a lysine residue of the acceptor substrate [1]. Assembly of a chain of >=4 ubiquitins linked together via Lys48 of ubiquitin marks cellular proteins for degradation by the 26S proteasome [2-3]. The 26S proteasome is a 2.5 megadalton macromolecular protein complex that comprises two distinct subparticles: the 19S cap regulatory particle (RP) and the 20S core
- Dose response confirmation of uHTS antagonist hits from Gli-SUFU in a luminescent reporter assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network(MLPCN) Grant Number: 1R03MH094195-01 Assay Provider: James Chen, Ph.D.,Stanford University, Stanford California The Hh pathway plays a critical role in the patterning of certain embryonic tissues and contributes to their neoplastic transformation later in life. Hh signaling regulates cerebellar patterning by promoting the proliferation of neuronal precursor cells, and constitutive Hh target gene expression can lead to medulloblastoma, the most common pediatric brain tumor(1). Hh signaling is normally initiated by the binding of Hh ligands to the twelve-pass transmembrane protein Ptch1(2) inducing its exit from the cilium, leading to Smo accumulation and activation within the antenna-like organelle. Activated Smo then shifts the balance between repressor and activator forms
- Dose response confirmation of uHTS hits for Apaf-1 in a Fluorescent assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBIMR, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: R01 CA136513 Assay Provider: Dr. Xuejun Jiang, Sloan-Kettering Institute for Cancer Research, New York, NY Apoptosis is a major form of programmed cell death that multicellular organisms utilize to maintain tissue homeostasis and to eliminate unwanted or damaged cells. It plays a critical role in development, immune responses and many other physiological events. In mammals, the mitochondrial cytochrome c-mediated apoptotic pathway is initiated by cytochrome c release from mitochondria. Deregulation of the cytochrome c apoptotic pathway can lead to diseases such as cancer, immune disorders, and neurodegenerative diseases. The overall goal of this project is to identify Apaf-1 inhibitors by high throughput screening. Apaf-1 is the essential mediato
- Dose response confirmation of uHTS hits for Apaf-1 using a LZ-Caspase-9/Caspase-3 Fluorescent Selectivity assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBIMR, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: R01 CA136513 Assay Provider: Dr. Xuejun Jiang, Sloan-Kettering Institute for Cancer Research, New York, NY Apoptosis is a major form of programmed cell death that multicellular organisms utilize to maintain tissue homeostasis and to eliminate unwanted or damaged cells. It plays a critical role in development, immune responses and many other physiological events. In mammals, the mitochondrial cytochrome c-mediated apoptotic pathway is initiated by cytochrome c release from mitochondria. Deregulation of the cytochrome c apoptotic pathway can lead to diseases such as cancer, immune disorders, and neurodegenerative diseases. The overall goal of this project is to identify Apaf-1 inhibitors by high throughput screening. Apaf-1 is the essential mediato
- Dose response confirmation uHTS hits for MazEF TA System activators via a fluorescence-based single-stranded RNase assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 2R01 GM068385-06 Assay Provider: Dr. Paul Hergenrother, University of Illinois, Urbana, IL Bacterial resistance to antibiotics is a worldwide health crisis. Resistance typically occurs as a result of chromosomal mutation or acquisition of a mobile genetic element, such as a plasmid, that harbors resistance-mediating genes. One of the most common plasmid maintenance systems is the toxin-antitoxin (TA) postsegregational killing mechanism. In this mechanism, if a plasmid-free daughter cell arises, the labile antitoxin is degraded and the toxin induces cell death. TA genes are ubiquitous in clinical isolates of certain drug-resistant bacteria, and it has been postulated that compounds that disrupt the TA interaction could free the toxin to kill the bac
- Dose-response confirmation of microRNA-mediated mRNA deadenylation inhibitors by fluoresence polarization assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network(MLPCN) Grant Number: R03 MH094198-01 Assay Provider: Kalle Gehring, Ph.D., McGill University The regulation of gene expression is a mechanism that allows cells to respond to growth and proliferation stimuli, stress, and nutrient availability. It is managed at multiple levels: mRNA expression, mRNA translation initiation and mRNA decay (1). MicroRNAs (miRNAs) are endogenous small RNAs that post-transcriptionally regulate gene expression to control a wide range of biological processes including cell growth, division and differentiation, as well as metabolism and development. The poly(A) tail of mRNA is bound by several molecules of the poly(A)-binding protein (PABP), an abundant cytoplasmic protein in eukaryotes that promotes translation (2). PABPC1 is a multi-domain protein t
- Enzymatic assay The aim of this in vitro assay was to measure the inhibition of HCV NS3/4A protease complexes by the compounds of the present invention. This assay provides an indication of how effective compounds of the present invention would be in inhibiting HCV NS3/4A proteolytic activity. The inhibition of full-length hepatitis C NS3 protease enzyme was measured essentially as described in Poliakov, 2002 Prot Expression & Purification 25 363 371. Briefly, the hydrolysis of a depsipeptide substrate, Ac-DED(Edans)EEAbu-y-[COO]ASK(Dabcyl)-NH2 (AnaSpec, San Jose, USA), was measured spectrofluorometrically in the presence of a peptide cofactor, KKGSVVIVGRIVLSGK (Ake Engstrom, Department of Medical Biochemistry and Microbiology, Uppsala University, Sweden). [Landro, 1997 #Biochem 36 9340-9348]. The enzyme (1 nM) was incubated in 50 mM HEPES, pH 7.5, 10 mM DTT, 40% glycerol, 0.1% n-octyl-D-glucoside, with 25 uM NS4A cofactor and inhibitor at 30 C. for 10 min.
- Estrogen Receptor-alpha Coactivator Binding Potentiators Dose Response Confirmation NIH Molecular Libraries Screening Centers Network [MLSCN] Emory Chemical Biology Discovery Center in MLSCN Assay provider: John A. Katzenellenbogen, University of Illinois Urbana-Champaign MLSCN Grant: 1 X01MH78953-01 Assay Overview Menopause is associated with the onset of hot flashes, night sweats, mood changes, and urogenital atrophy, which many women find distressing enough to seek medical management for relief. Estrogens in the form of hormone therapy (HT) have been the standard treatment for menopausal symptoms for decades. Although HT is the most effective treatment for hot flashes, the Women's Health Initiative (WHI) trial found that the combination of estrogen and progestin increases a woman's risk for heart disease, stroke, breast cancer, venous thromboembolic events, and dementia and does not improve quality of life indices such as emotional and sexual functioning and vitality. The adverse effects of HT created a large unmet need for selective estrogen rece
- Fluorescent Polarization Homogeneous Dose Response HTS to Indentify Inhibitors of Mex-5 Binding to TCR-2 Broad Institute: MLPCN maternal gene expression Project ID: 2024 Keywords: Zinc finger, C. elegans, maternal gene expression, RNA-protein interaction, gene regulation, MEX-5, POS-1, embryonic development Primary Collaborators: Sean Ryder, U. Mass Medical School, sean.ryder@umassmed.edu Project Overview: In most organisms, the body axes and founding tissue types are formed prior to the onset of zygotic transcription. Thus, post-transcriptional regulation of maternal transcripts by maternally supplied RNA-binding proteins is crucial to early patterning events. Few tools exist to study specific regulatory networks guided by RNA-binding proteins during early development. Importantly, standard genetic analyses are complicated by the maternal effect, pleiotropy, and embryonic lethality. This screen aims to identify small molecule inhibitors of RNA-binding protein function for two proteins required for Caenorhabditis elegans early development. The rationale was two fold. First, inhib
- Fluorescent Polarization Homogeneous Dose Response HTS to Indentify Inhibitors of POS-1 Binding to mex-3-RNA Broad Institute: MLPCN maternal gene expression Project ID: 2024 Keywords: Zinc finger, C. elegans, maternal gene expression, RNA-protein interaction, gene regulation, MEX-5, POS-1, embryonic development Primary Collaborators: Sean Ryder, U. Mass Medical School, sean.ryder@umassmed.edu Project Overview: In most organisms, the body axes and founding tissue types are formed prior to the onset of zygotic transcription. Thus, post-transcriptional regulation of maternal transcripts by maternally supplied RNA-binding proteins is crucial to early patterning events. Few tools exist to study specific regulatory networks guided by RNA-binding proteins during early development. Importantly, standard genetic analyses are complicated by the maternal effect, pleiotropy, and embryonic lethality. This screen aims to identify small molecule inhibitors of RNA-binding protein function for two proteins required for Caenorhabditis elegans early development. The rationale was two fold. First, inhib
- HTS Dose response counterscreen for assays utilizing the enzyme, b-galactosidase Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak , Duke University, Durham NC b-galactosidase (b-gal), a hydrolase enzyme that catalyzes the hydrolysis of b-galactosides to monosaccharides is utilized in many different screening technologies involving enzyme reaction coupling and reporter assays, for example DiscoverX b-Arrestin GPCR assays such as the KOR1 Agonist or Antagonist. This assay was developed and performed as a counterscreen for screening assays that utilize b-gal and a reaction that it catalyzes. By detecting inhibitors and activators of this enzyme, it is possible to attribute activity not to the primary assay in question, but rather to interaction with the method of detection. References Fowler et al. (1970). "The amino acid sequ
- HTS Dose response counterscreen for assays utilizing the enzyme, beta-galactosidase - Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak , Duke University, Durham, NC b-galactosidase (b-gal), a hydrolase enzyme that catalyzes the hydrolysis of b-galactosides to monosaccharides is utilized in many different screening technologies involving enzyme reaction coupling and reporter assays, for example DiscoverX b-Arrestin GPCR assays such as the APJ Agonist or Antagonist. This assay was developed and performed as a counterscreen for screening assays that utilize b-gal and a reaction that it catalyzes. By detecting inhibitors and activators of this enzyme, it is possible to attribute activity not to the primary assay in question, but rather to interaction with the method of detection. References Fowler et al. (1970). "The amino aci
- HTS Dose response counterscreen for assays utilizing the enzyme, beta-galactosidase - Set 3 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak , Duke University, Durham, NC b-galactosidase (b-gal), a hydrolase enzyme that catalyzes the hydrolysis of b-galactosides to monosaccharides is utilized in many different screening technologies involving enzyme reaction coupling and reporter assays. This assay was developed and performed as a counterscreen for the primary assay originally identified as "uHTS identification of small molecule antagonists of the CCR6 receptor via a luminescent beta-arrestin assay", AID 493098. By detecting inhibitors and activators of this enzyme, it is possible to attribute activity not to the primary assay in question, but rather to interaction with the method of detection. References Fowler et al. (1970).
- HTS Image-Based Screen for Agonists of the DOR Receptor Data Source: Sanford-Burnham Center for Chemical Genomics Source Affiliation: Sanford-Burnham Medical Research Institute Network: NIH Molecular Libraries Probe Production Centers Network Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak, Duke University, Durham, NC Drug addiction is a disease that involves the G-protein coupled receptors in the central nervous system resulting in compulsive or abnormal behavior. Recent studies have shown that opioid receptors play a role in regulating other receptors that interact with drug and other substance abuse. The opioid receptors are composed of multiple of multiple receptor subtypes whose contribution the addictive behaviors are not fully delineated. The aim of this assay is to identify small molecule agonists of the human delta opioid receptor (DOR). The dose response assay is developed and performed to evaluate selectivity of hits originally identified in a uHTS luminescent beta-arrestin assay for agonists of the KOR re
- HTS Image-Based Screen for Antagonists of the DOR Receptor Data Source: Sanford-Burnham Center for Chemical Genomics Source Affiliation: Sanford-Burnham Medical Research Institute Network: NIH Molecular Libraries Probe Production Centers Network Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak, Duke University, Durham, NC Drug addiction is a disease that involves the G-protein coupled receptors in the central nervous system resulting in compulsive or abnormal behavior. Recent studies have shown that opioid receptors play a role in regulating other receptors that interact with drug and other substance abuse. The opioid receptors are composed of multiple of multiple receptor subtypes whose contribution the addictive behaviors are not fully delineated. The aim of this assay is to identify small molecule antagonists of the human delta opioid receptor (DOR). The dose response assay is developed and performed to evaluate selectivity of hits originally identified in a uHTS luminescent beta-arrestin assay for agonists of the KOR
- High Throughput Fluorescence Polarization Screen for Bcl-B Phenotype Converters Sanford-Burnham Center for Chemical Genomics (SBCCG) Sanford-Burnham Medical Research Institute (San Diego, CA) NIH Molecular Libraries Screening Centers Network (MLSCN) Bcl-B is an anti-apoptotic member of the Bcl-2 family that is prominently expressed in plasma and multiple myeloma cells. TR3 (NR4A1; HMR; NP10; GFRP1; NAK1; NUR77; NGFIB) is an orphan member of the steroid/thyroid/retinoid nuclear receptor superfamily that translocates from cellular nuclei to mitochondria upon exposure to various pro-apoptotic stimuli. At mitochondria, TR3 binds to Bcl-B and converts it into a pro-apoptotic protein. A specific 9-amino acid sequence within the full length TR3 is able to form a complex with Bcl-B and induce its phenotypic conversion. This interaction forms the basis for a high throughput fluorescence polarization screen for compound mimetics. Such compounds may be useful as chemical probes for understanding the functional mechanism of Bcl-B, and may have therapeutic relevance in a
- Human Glucose-6-Phosphate Dehydrogenase Dose Response Selectivity Assay for Inhibitors of Plasmodium falciparum Glucose-6-Phosphate Dehydrogenase Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1R21AI082434-01 Assay Provider: Lars Bode, Ph.D., University of California San Diego, San Diego, CA Tropical malaria caused by the protozoan parasite Plasmodium falciparum is responsible for up to three million deaths annually. Due to increasing regional distribution and resistances against the clinically used antimalarials, novel antimalarial drugs - which have new mechanisms of action and are suitable for combination therapies - are urgently required. Plasmodium falciparum Glucose-6-phosphate dehydrogenase (PfGluPho) is a potential novel target for antimalarial drug design. Glucose-6-Phosphate Dehydrogenase (G6PD) reaction is the first and rate-limiting step in the pentose phosphate pathway (PPP), catalyzed by a bifunctional enzyme Plasmodium fal
- Image-Based HTS for Selective Antagonists of GPR35 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Proposal Number: 1X01MH085708-01 Assay Provider: Dr. Lawrence Barak, Duke University Addictive behavior stems from abnormal signaling activities in the brain. Thus identification of compounds blocking this modified signaling activity may lead to treatments for addictive behavior. GPR35, a to-date uncharacterized orphan G-Protein Coupled Receptor, is thought to play a role in addiction and has homology to other known receptors of abuse. The goal of this project is to identify small molecule antagonists of GPR35, which may aid in characterization of this receptor and ultimately further the understanding of the role of GPR35 in addictive behavior. This high content imaging assay utilizes a cell line permanently expressing a beta-arrestin GFP biosensor and th
- In Vitro MKP-3 Dose Response Assay for SAR Study Data Source: Burnham Center for Chemical Genomics (BCCG) Source Affiliation: Burnham Institute for Medical Research (BIMR, La Jolla, CA) Network: NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Number: XO1 MH076390-01 Assay Provider: Dr. John Lazo, University of Pittsburg This MKP-3 dose response assay is developed and performed for the purpose of SAR study on analogs of hits originally identified in the MKP-3 in vitro HTS assay (AID 425) MKP-3 (mitogen-activated protein kinase phosphatase-3; EC 3.1.3.48, EC 3.1.3.16), a dual specificity phosphatase negatively regulates ERK1/2 by catalyzing the removal of a phosphoryl group from Thr(P) and Tyr(P) in the activation loop consensus motif -pTXpY. MKP-3 screening was performed using a biochemical assay developed at the laboratory of Prof. John Lazo (University of Pittsburg). The assay was optimized and run at the Burnham Center for Chemical Genomics (BCCG) as part of the Molecular Library Screening Center Network (MLSCN
- Inhibition of rhGAA Determined by Measuring the Residual Activity in the Presence of an Inhibitor in Vitro The rhGAA enzyme sold under the name Myozyme used comes from residues of perfusions of the recombinant enzyme used for treating, by enzyme therapy, the patients with Pompe disease in the Department of Translational Medical Sciences, University of Naples Federico II (IT). The compounds are solubilized at various concentrations in a 100 mM sodium acetate buffer, pH 4.0, as well as the 4-nitrophenyl-α-D-glucopyranoside substrate (20 mM). After 2 minutes of equilibration of the temperature at 37 C., the rhGAA enzyme in solution in the same buffer is added (total volume: 200 μL). After 2 minutes of reaction at 37 C., a 1M solution of sodium carbonate (800 μL), pH 11.0, is added and the mixture is cooled in ice. The absorbance of the solution is measured at 420 nm at ambient temperature.
- Inhibitors of Plasmodium falciparum M1- Family Alanyl Aminopeptidase (M1AAP) Southern Research Molecular Libraries Screening Center (SRMLSC) Southern Research Institute (Birmingham, Alabama) NIH Molecular Libraries Screening Centers Network (MLSCN) Assay Provider: Dr. Donald Gardiner, Queensland Institute of Medical Research Grant: 1-R03-MH082342-01A1 The intraerythrocytic stages of the human malaria parasite Plasmodium falciparum employs two cytosolic neutral aminopeptidases, an M1-family alanyl aminopeptidase (M1AAP) and an M17-family leucine aminopeptidase (M17LAP), in the terminal stages of host hemoglobin digestion. Their action results in the release of free amino acids that are used for the anabolism of parasite proteins and, hence, are critical to the development of the parasite in red blood cells. Inhibitors of the two exopeptidases prevent the growth of P. falciparum parasites in vitro, and protect mice from infection with rodent malaria P. chabaudi, providing strong evidence that these enzymes are targets which can be used to develop new anti-ma
- Inhibitors of Plasmodium falciparum M17- Family Leucine Aminopeptidase (M17LAP) Southern Research Molecular Libraries Screening Center (SRMLSC) Southern Research Institute (Birmingham, Alabama) NIH Molecular Libraries Screening Centers Network (MLSCN) Assay Provider: Dr. Donald Gardiner, Queensland Institute of Medical Research Award: 1-R03-MH082342-01A1 The intraerythrocytic stages of the human malaria parasite Plasmodium falciparum employs two cytosolic neutral aminopeptidases, an M1-family alanyl aminopeptidase (M1AAP) and an M17-family leucine aminopeptidase (M17LAP), in the terminal stages of host hemoglobin digestion. Their action results in the release of free amino acids that are used for the anabolism of parasite proteins and, hence, are critical to the development of the parasite in red blood cells. Inhibitors of the two exopeptidases prevent the growth of P. falciparum parasites in vitro, and protect mice from infection with rodent malaria P. chabaudi, providing strong evidence that these enzymes are targets which can be used to develop new anti-ma
- LYP1 Fluorescent Assay using OMFP substrate for In Vitro dose response studies. Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1R21NS056945-01 Assay Provider: Dr. Nunzio Bottini, La Jolla Institute for Allergy and Immunology, La Jolla, CA LYP, is a lymphocyte specific protein tyrosine phosphatase that plays a critical regulatory role in T cell receptor signaling. The PTPN22 gene encodes this phosphatase. A single-nucleotide polymorphism in PTPN22 is associated with a number of autoimmune disorders, including type 1 diabetes, rheumatoid arthritis, juvenile rheumatoid arthritis, systemic lupus erythematosus and Grave's disease. The autoimmunity-predisposing allele is a gain-of-function mutant suggesting that a specific small-molecule inhibitor could eliminate its effect. This biochemical assay employs a fluorescent readout based on the enzyme's ability to liberate
- Luminescent assay for identification of inhibitors of human intestinal alkaline phosphatase Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Proposal Number: XO1 MH077602-01 Alkaline phosphatase (EC 3.1.3.1) (APs) catalyze the hydrolysis of phosphomonoesters, releasing inorganic phosphate and alcohol. APs are dimeric enzymes found in most organisms. In human, four isozymes of APs have been identified. One isozyme is tissue-nonspecific (designated TNAP) and three other isozymes are tissue-specific and named according to the tissue of their predominant expression: intestinal (IAP), placental (PLAP) and germ cell (GCAP) alkaline phosphatases. IAP expression is largely restricted to the gut, especially to the epithelial cells (enterocytes) of the small intestinal mucosa. IAP is inhibited by a number of inhibitors (1). They include L-phenylalanine, (2, 3), L-tryptophan (4), L-leucine and phenylalanine-g
- MKP-3 in vitro HTS assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Number: XO1 MH076390-01 Assay Provider: Dr. John Lazo, University of Pittsburg MKP-3 (mitogen-activated protein kinase phosphatase-3; EC 3.1.3.48, EC 3.1.3.16), a dual specificity phosphatase negatively regulates ERK1/2 by catalyzing the removal of a phosphoryl group from Thr(P) and Tyr(P) in the activation loop consensus motif -pTXpY. MKP-3 screening was performed using a biochemical assay developed at the laboratory of Prof. John Lazo (University of Pittsburg). The assay was optimized and run at the Sanford-Burnham Center for Chemical Genomics (SBCCG) as part of the Molecular Library Screening Center Network (MLSCN). Enzyme activity and its inhibition by screened compounds was measured in an end-point assay based on hydrolysis of 3-O-methylfluorescein phosphate
- SAR Analysis of Agonists of the DOR Receptor using an Image-Based Assay - Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network(MLPCN) Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak, Duke University, Durham NC Drug addiction is a disease that involves the G-protein coupled receptors in the central nervous system resulting in compulsive or abnormal behavior. Recent studies have shown that opioid receptors play a role in regulating other receptors that interact with drug and other substance abuse. The opioid receptors are composed of multiple of multiple receptor subtypes whose contribution the addictive behaviors are not fully delineated. The aim of this assay is to identify small molecule agonists of the human delta opioid receptor (DOR). This dose response assay is developed and performed to confirm hits originally identified in "uHTS identification of small molecule agonists of
- SAR Analysis of Agonists of the DOR Receptor using an Image-Based Assay - Set 3 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network(MLPCN) Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak, Duke University, Durham NC Drug addiction is a disease that involves the G-protein coupled receptors in the central nervous system resulting in compulsive or abnormal behavior. Recent studies have shown that opioid receptors play a role in regulating other receptors that interact with drug and other substance abuse. The opioid receptors are composed of multiple of multiple receptor subtypes whose contribution the addictive behaviors are not fully delineated. The aim of this assay is to identify small molecule agonists of the human delta opioid receptor (DOR). This dose response assay is developed and performed to confirm hits originally identified in "uHTS identification of small molecule agonists of
- SAR Analysis of Agonists of the DOR Receptor using an Image-Based Assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network(MLPCN) Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak, Duke University, Durham NC Drug addiction is a disease that involves the G-protein coupled receptors in the central nervous system resulting in compulsive or abnormal behavior. Recent studies have shown that opioid receptors play a role in regulating other receptors that interact with drug and other substance abuse. The opioid receptors are composed of multiple of multiple receptor subtypes whose contribution the addictive behaviors are not fully delineated. The aim of this assay is to identify small molecule agonists of the human delta opioid receptor (DOR). This dose response assay is developed and performed to confirm hits originally identified in "uHTS identification of small molecule agonists of
- SAR Analysis of Agonists of the MOR Receptor using an Image-Based Assay - Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network(MLPCN) Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak, Duke University, Durham NC Drug addiction is a disease that involves the G-protein coupled receptors in the central nervous system resulting in compulsive or abnormal behavior. Recent studies have shown that opioid receptors play a role in regulating other receptors that interact with drug and other substance abuse. The opioid receptors are composed of multiple of multiple receptor subtypes whose contribution the addictive behaviors are not fully delineated. The aim of this assay is to identify small molecule agonists of the human mu opioid receptor (MOR). This dose response assay is developed and performed to confirm hits originally identified in "uHTS identification of small molecule agonists of th
- SAR Analysis of Agonists of the MOR Receptor using an Image-Based Assay - Set 3 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network(MLPCN) Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak, Duke University, Durham NC Drug addiction is a disease that involves the G-protein coupled receptors in the central nervous system resulting in compulsive or abnormal behavior. Recent studies have shown that opioid receptors play a role in regulating other receptors that interact with drug and other substance abuse. The opioid receptors are composed of multiple of multiple receptor subtypes whose contribution the addictive behaviors are not fully delineated. The aim of this assay is to identify small molecule agonists of the human mu opioid receptor (MOR). This dose response assay is developed and performed to confirm hits originally identified in "uHTS identification of small molecule agonists of th
- SAR Analysis of Agonists of the MOR Receptor using an Image-Based Assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network(MLPCN) Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak, Duke University, Durham NC Drug addiction is a disease that involves the G-protein coupled receptors in the central nervous system resulting in compulsive or abnormal behavior. Recent studies have shown that opioid receptors play a role in regulating other receptors that interact with drug and other substance abuse. The opioid receptors are composed of multiple of multiple receptor subtypes whose contribution the addictive behaviors are not fully delineated. The aim of this assay is to identify small molecule agonists of the human mu opioid receptor (MOR). This dose response assay is developed and performed to confirm hits originally identified in "uHTS identification of small molecule agonists of th
- SAR Analysis of Antagonists of the DOR Receptor using an Image-Based Assay - Set 3 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network(MLPCN) Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak, Duke University, Durham NC Drug addiction is a disease that involves the G-protein coupled receptors in the central nervous system resulting in compulsive or abnormal behavior. Recent studies have shown that opioid receptors play a role in regulating other receptors that interact with drug and other substance abuse. The opioid receptors are composed of multiple of multiple receptor subtypes whose contribution the addictive behaviors are not fully delineated. The aim of this assay is to identify small molecule antagonists of the human delta opioid receptor (DOR). This dose response assay is developed and performed to confirm hits originally identified in "uHTS identification of small molecule ago
- SAR Analysis of Antagonists of the DOR Receptor using an Image-Based Assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network(MLPCN) Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak, Duke University, Durham NC Drug addiction is a disease that involves the G-protein coupled receptors in the central nervous system resulting in compulsive or abnormal behavior. Recent studies have shown that opioid receptors play a role in regulating other receptors that interact with drug and other substance abuse. The opioid receptors are composed of multiple of multiple receptor subtypes whose contribution the addictive behaviors are not fully delineated. The aim of this assay is to identify small molecule antagonists of the human delta opioid receptor (DOR). This dose response assay is developed and performed to confirm hits originally identified in "uHTS identification of small molecule agonists
- SAR Analysis of Antagonists of the MOR Receptor using an Image-Based Assay - Set 3 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network(MLPCN) Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak, Duke University, Durham NC Drug addiction is a disease that involves the G-protein coupled receptors in the central nervous system resulting in compulsive or abnormal behavior. Recent studies have shown that opioid receptors play a role in regulating other receptors that interact with drug and other substance abuse. The opioid receptors are composed of multiple of multiple receptor subtypes whose contribution the addictive behaviors are not fully delineated. The aim of this assay is to identify small molecule antagonists of the human mu opioid receptor (MOR). This dose response assay is developed and performed to confirm hits originally identified in "uHTS identification of small molecule agoni
- SAR Analysis of Antagonists of the MOR Receptor using an Image-Based Assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network(MLPCN) Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak, Duke University, Durham NC Drug addiction is a disease that involves the G-protein coupled receptors in the central nervous system resulting in compulsive or abnormal behavior. Recent studies have shown that opioid receptors play a role in regulating other receptors that interact with drug and other substance abuse. The opioid receptors are composed of multiple of multiple receptor subtypes whose contribution the addictive behaviors are not fully delineated. The aim of this assay is to identify small molecule antagonists of the human mu opioid receptor (MOR). This dose response assay is developed and performed to confirm hits originally identified in "uHTS identification of small molecule agonists of
- SAR Analysis of Selective Antagonists of GPR55 using an Image-Based Assay Data Source: Sanford-Burnham Center for Chemical Genomics(SSBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1X01 MH085708-01 Assay Provider: Dr. Lawrence Barak, Duke University, Durham, NC Addictive behavior stems from abnormal signaling activities in the brain. Thus identification of compounds blocking this modified signaling activity may lead to treatments for addictive behavior. GPR35, a to-date uncharacterized orphan G-Protein Coupled Receptor, is thought to play a role in addiction and has homology to other known receptors of abuse. This high-content imaging assay was used as a counter screen for hits originally identified in a high-content screen for antagonists of the GPR35 receptor "Image-based HTS for Selective Antagonists of GPR35" (AID 2058) and to study the structure-activity relationship on analogs of the confirmed hits. Compounds are ei
- SAR LYP1 Fluorescent Assay using OMFP substrate for In Vitro dose response studies - Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1R21NS056945-01 Assay Provider: Dr. Nunzio Bottini, La Jolla Institute for Allergy and Immunology CA LYP, is a lymphocyte specific protein tyrosine phosphatase that plays a critical regulatory role in T cell receptor signaling. The PTPN22 gene encodes this phosphatase. A single-nucleotide polymorphism in PTPN22 is associated with a number of autoimmune disorders, including type 1 diabetes, rheumatoid arthritis, juvenile rheumatoid arthritis, systemic lupus erythematosus and Grave's disease. The autoimmunity-predisposing allele is a gain-of-function mutant suggesting that a specific small-molecule inhibitor could eliminate its effect. This biochemical assay employs a fluorescent readout based on the enzyme's ability to liberate methyl-fluo
- SAR LYP1 Fluorescent Assay using OMFP substrate for In Vitro dose response studies - Set 3 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1R21NS056945-01 Assay Provider: Dr. Nunzio Bottini, La Jolla Institute for Allergy and Immunology, La Jolla, CA LYP, is a lymphocyte specific protein tyrosine phosphatase that plays a critical regulatory role in T cell receptor signaling. The PTPN22 gene encodes this phosphatase. A single-nucleotide polymorphism in PTPN22 is associated with a number of autoimmune disorders, including type 1 diabetes, rheumatoid arthritis, juvenile rheumatoid arthritis, systemic lupus erythematosus and Grave's disease. The autoimmunity-predisposing allele is a gain-of-function mutant suggesting that a specific small-molecule inhibitor could eliminate its effect. This biochemical assay employs a fluorescent readout based on the enzyme's ability to liberate
- SAR LYP1 Fluorescent Assay using OMFP substrate for In Vitro dose response studies - Set 4 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1R21NS056945-01 Assay Provider: Dr. Nunzio Bottini , La Jolla Institute for Allergy and Immunology CA LYP, is a lymphocyte specific protein tyrosine phosphatase that plays a critical regulatory role in T cell receptor signaling. The PTPN22 gene encodes this phosphatase. A single-nucleotide polymorphism in PTPN22 is associated with a number of autoimmune disorders, including type 1 diabetes, rheumatoid arthritis, juvenile rheumatoid arthritis, systemic lupus erythematosus and Grave's disease. The autoimmunity-predisposing allele is a gain-of-function mutant suggesting that a specific small-molecule inhibitor could eliminate its effect. This biochemical assay employs a fluorescent readout based on the enzyme's ability to liberate methyl-fl
- SAR LYP1 Fluorescent Assay using OMFP substrate for In Vitro dose response studies Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1R21NS056945-01 Assay Provider: Dr. Nunzio Bottini, La Jolla Institute for Allergy and Immunology, La Jolla, CA LYP, is a lymphocyte specific protein tyrosine phosphatase that plays a critical regulatory role in T cell receptor signaling. The PTPN22 gene encodes this phosphatase. A single-nucleotide polymorphism in PTPN22 is associated with a number of autoimmune disorders, including type 1 diabetes, rheumatoid arthritis, juvenile rheumatoid arthritis, systemic lupus erythematosus and Grave's disease. The autoimmunity-predisposing allele is a gain-of-function mutant suggesting that a specific small-molecule inhibitor could eliminate its effect. This biochemical assay employs a fluorescent readout based on the enzyme's ability to liberate
- SAR LYP1 Fluorescent Assay using pCAP substrate for In Vitro dose response studies - Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1R21NS056945-01 Assay Provider: Dr. Nunzio Bottini, La Jolla Institute for Allergy and Immunology LYP, is a lymphocyte specific protein tyrosine phosphatase that plays a critical regulatory role in T cell receptor signaling. The PTPN22 gene encodes this phosphatase. A single-nucleotide polymorphism in PTPN22 is associated with a number of autoimmune disorders, including type 1 diabetes, rheumatoid arthritis, juvenile rheumatoid arthritis, systemic lupus erythematosus and Grave's disease. The autoimmunity-predisposing allele is a gain-of-function mutant suggesting that a specific small-molecule inhibitor could eliminate its effect. Finding specific inhibitors of protein phosphatases has proven extremely difficult. The goal of this project is to
- SAR LYP1 Fluorescent Assay using pCAP substrate for In Vitro dose response studies - Set 3 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1R21NS056945-01 Assay Provider: Dr. Nunzio Bottini, La Jolla Institute for Allergy and Immunology, La Jolla, CA LYP, is a lymphocyte specific protein tyrosine phosphatase that plays a critical regulatory role in T cell receptor signaling. The PTPN22 gene encodes this phosphatase. A single-nucleotide polymorphism in PTPN22 is associated with a number of autoimmune disorders, including type 1 diabetes, rheumatoid arthritis, juvenile rheumatoid arthritis, systemic lupus erythematosus and Grave's disease. The autoimmunity-predisposing allele is a gain-of-function mutant suggesting that a specific small-molecule inhibitor could eliminate its effect. Finding specific inhibitors of protein phosphatases has proven extremely difficult. The goal of this
- SAR LYP1 Fluorescent Assay using pCAP substrate for In Vitro dose response studies - Set 4 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network(MLPCN) Grant Number: 1R21NS056945-01 Assay Provider: Dr. Nunzio Bottini, La Jolla Institute for Allergy and Immunology, La Jolla, CA LYP, is a lymphocyte specific protein tyrosine phosphatase that plays a critical regulatory role in T cell receptor signaling. The PTPN22 gene encodes this phosphatase. A single-nucleotide polymorphism in PTPN22 is associated with a number of autoimmune disorders, including type 1 diabetes, rheumatoid arthritis, juvenile rheumatoid arthritis, systemic lupus erythematosus and Grave's disease. The autoimmunity-predisposing allele is a gain-of-function mutant suggesting that a specific small-molecule inhibitor could eliminate its effect. Finding specific inhibitors of protein phosphatases has proven extremely difficult. The goal of this
- SAR LYP1 Fluorescent Assay using pCAP substrate for In Vitro dose response studies Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1R21NS056945-01 Assay Provider: Dr. Nunzio Bottini, La Jolla Institute for Allergy and Immunology, La Jolla, CA LYP, is a lymphocyte specific protein tyrosine phosphatase that plays a critical regulatory role in T cell receptor signaling. The PTPN22 gene encodes this phosphatase. A single-nucleotide polymorphism in PTPN22 is associated with a number of autoimmune disorders, including type 1 diabetes, rheumatoid arthritis, juvenile rheumatoid arthritis, systemic lupus erythematosus and Grave's disease. The autoimmunity-predisposing allele is a gain-of-function mutant suggesting that a specific small-molecule inhibitor could eliminate its effect. Finding specific inhibitors of protein phosphatases has proven extremely difficult. The goal of thi
- SAR Selectivity Analysis of small molecule inhibitors of PEST using pCAP in a fluorescence assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network(MLPCN) Grant Number: 1R21NS056945-01 Assay Provider: Dr. Nunzio Bottini, La Jolla Institute for Allergy and Immunology, La Jolla, CA PTP-PEST, encoded by PTPN12 gene, is a protein tyrosine phosphatase that contains a C-terminal PEST motif, which serves as a protein-protein interaction domain. This PTP dephosphorylates c-ABL, and may play a role in oncogenesis. This is a selectivity counterscreen to aid with identification of specific inhibitors of LYP phosphatase (AID 2135). This assay uses a phosphorylated coumarin amino acid (pCAP) moiety attached to a 14-mer peptide, which is a substrate for LYP. This dose response assay is developed and performed to assess selectivity of hits originally found in "uHTS identification of small molecule inhibitors of LYP via a fluo
- SAR analysis of Agonists of the Kappa Opioid Receptor (KOR) using an Image-Based Assay - Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics Source Affiliation: Sanford-Burnham Medical Research Institute Network: NIH Molecular Libraries Production Centers Network Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak, Duke University, Durham NC Drug addiction is a disease that involves the G-protein coupled receptors in the central nervous system resulting in compulsive or abnormal behavior. Recent studies have shown that opioid receptors play a role in regulating other receptors that interact with drug and other substance abuse. The opioid receptors are composed of multiple of multiple receptor subtypes whose contribution the addictive behaviors are not fully delineated. The aim of this assay is to identify small molecule agonists of the human kappa opioid receptor (KOR). This dose response assay is developed and performed to confirm hits originally identified in "uHTS identification of small molecule agonists of the kappa opioid receptor via a lum
- SAR analysis of Agonists of the Kappa Opioid Receptor (KOR) using an Image-Based Assay Data Source: Sanford-Burnham Center for Chemical Genomics Source Affiliation: Sanford-Burnham Medical Research Institute Network: NIH Molecular Libraries Production Centers Network Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak, Duke University, Durham NC Drug addiction is a disease that involves the G-protein coupled receptors in the central nervous system resulting in compulsive or abnormal behavior. Recent studies have shown that opioid receptors play a role in regulating other receptors that interact with drug and other substance abuse. The opioid receptors are composed of multiple of multiple receptor subtypes whose contribution the addictive behaviors are not fully delineated. The aim of this assay is to identify small molecule agonists of the human kappa opioid receptor (KOR). This dose response assay is developed and performed to confirm hits originally identified in "uHTS identification of small molecule agonists of the kappa opioid receptor via a lum
- SAR analysis of Agonists of the Kappa Opioid Receptor (KOR) using an Image-Based Assay-Set 3 Data Source: Sanford-Burnham Center for Chemical Genomics Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak, Duke University, Durham NC Drug addiction is a disease that involves the G-protein coupled receptors in the central nervous system resulting in compulsive or abnormal behavior. Recent studies have shown that opioid receptors play a role in regulating other receptors that interact with drug and other substance abuse. The opioid receptors are composed of multiple of multiple receptor subtypes whose contribution the addictive behaviors are not fully delineated. The aim of this assay is to identify small molecule agonists of the human kappa opioid receptor (KOR). This dose response assay is developed and performed to confirm hits originally identified in "uHTS identification of small molecule agonists of the kappa op
- SAR analysis of Antagonists of the GPR35 Receptor using an Image-Based Assay - Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics Source Affiliation: Sanford-Burnham Medical Research Institute Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1X01MH085708-01 Assay Provider: Dr. Lawrence Barak, Duke University Addictive behavior stems from abnormal signaling activities in the brain. Thus identification of compounds blocking this modified signaling activity may lead to treatments for addictive behavior. GPR35, a to-date uncharacterized orphan G-Protein Coupled Receptor, is thought to play a role in addiction and has homology to other known receptors of abuse. This high-content imaging assay is developed and performed to confirm activity of hits originally identified in a high-content screen for antagonists of the GPR35 receptor, "Image-Based HTS for Selective Antagonists of GPR35" (AID 2058) and to study the structure-activity relationship on analogs of the confirmed hits. Compounds are either acquired from commercial
- SAR analysis of Antagonists of the Kappa Opioid Receptor (KOR) using an Image-Based Assay - Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics Source Affiliation: Sanford-Burnham Medical Research Institute Network: NIH Molecular Libraries Production Centers Network Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak, Duke University, Durham NC Drug addiction is a disease that involves the G-protein coupled receptors in the central nervous system resulting in compulsive or abnormal behavior. Recent studies have shown that opioid receptors play a role in regulating other receptors that interact with drug and other substance abuse. The opioid receptors are composed of multiple of multiple receptor subtypes whose contribution the addictive behaviors are not fully delineated. The aim of this assay is to identify small molecule antagonists of the human kappa opioid receptor (KOR). This dose response assay is developed and performed to confirm hits originally identified in "uHTS identification of small molecule agonists of the kappa opioid receptor via a
- SAR analysis of Antagonists of the Kappa Opioid Receptor (KOR) using an Image-Based Assay - Set 3 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak, Duke University, Durham NC Drug addiction is a disease that involves the G-protein coupled receptors in the central nervous system resulting in compulsive or abnormal behavior. Recent studies have shown that opioid receptors play a role in regulating other receptors that interact with drug and other substance abuse. The opioid receptors are composed of multiple of multiple receptor subtypes whose contribution the addictive behaviors are not fully delineated. The aim of this assay is to identify small molecule antagonists of the human kappa opioid receptor (KOR). This dose response assay is developed and performed to confirm hits originally identified in "uHTS identification of small molecule agonists
- SAR analysis of Antagonists of the Kappa Opioid Receptor (KOR) using an Image-Based Assay - Set 4 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak, Duke University, Durham NC Drug addiction is a disease that involves the G-protein coupled receptors in the central nervous system resulting in compulsive or abnormal behavior. Recent studies have shown that opioid receptors play a role in regulating other receptors that interact with drug and other substance abuse. The opioid receptors are composed of multiple of multiple receptor subtypes whose contribution the addictive behaviors are not fully delineated. The aim of this assay is to identify small molecule antagonists of the human kappa opioid receptor (KOR). This dose response assay is developed and performed to confirm hits originally identified in "uHTS identification of small molecule agonists
- SAR analysis of Antagonists of the Kappa Opioid Receptor (KOR) using an Image-Based Assay Data Source: Sanford-Burnham Center for Chemical Genomics Source Affiliation: Sanford-Burnham Medical Research Institute Network: NIH Molecular Libraries Production Centers Network Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak, Duke University, Durham NC Drug addiction is a disease that involves the G-protein coupled receptors in the central nervous system resulting in compulsive or abnormal behavior. Recent studies have shown that opioid receptors play a role in regulating other receptors that interact with drug and other substance abuse. The opioid receptors are composed of multiple of multiple receptor subtypes whose contribution the addictive behaviors are not fully delineated. The aim of this assay is to identify small molecule antagonists of the human kappa opioid receptor (KOR). This dose response assay is developed and performed to confirm hits originally identified in "uHTS identification of small molecule agonists of the kappa opioid receptor via a
- SAR analysis of compounds that inhibit Human Immunodeficiency Virus Fusion. Data Source: Burnham Center for Chemical Genomics (BCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1R21NS059403-01 Assay Provider: Dr. Miriam Gochin, Touro University-California, Vallejo, CA The fusion-active conformation of the envelope protein gp41 of HIV-1 consists of an N-terminal trimeric a-helical coiled coil domain, and three anti-parallel C-terminal helices which fold down the grooves of the coiled coil to form a six-helix bundle. Disruption of the six-helix bundle is considered to be a key component of an effective non-peptide fusion inhibitor. This structure forms as a result of a conformational change in gp41, triggered by gp120 and co-receptor binding to host cell receptors. Prevention of six-helix bundle formation has been recognized as an important mechanism for viral fusion inhibition A metallopeptide-based fluorescence assay has been develope
- SAR analysis of small molecule Activators of Apaf-1 in a Fluorescent assay - set 2 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBIMR, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: R01 CA136513 Assay Provider: Dr. Xuejun Jiang, Sloan-Kettering Institute for Cancer Research, New York, NY Apoptosis is a major form of programmed cell death that multicellular organisms utilize to maintain tissue homeostasis and to eliminate unwanted or damaged cells. It plays a critical role in development, immune responses and many other physiological events. In mammals, the mitochondrial cytochrome c-mediated apoptotic pathway is initiated by cytochrome c release from mitochondria. Deregulation of the cytochrome c apoptotic pathway can lead to diseases such as cancer, immune disorders, and neurodegenerative diseases. The overall goal of this project is to identify Apaf-1 activators by high throughput screening. Apaf-1 is the essential mediato
- SAR analysis of small molecule Inhibitors of Apaf-1 in a Fluorescent assay - set 2 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBIMR, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: R01 CA136513 Assay Provider: Dr. Xuejun Jiang, Sloan-Kettering Institute for Cancer Research, New York, NY Apoptosis is a major form of programmed cell death that multicellular organisms utilize to maintain tissue homeostasis and to eliminate unwanted or damaged cells. It plays a critical role in development, immune responses and many other physiological events. In mammals, the mitochondrial cytochrome c-mediated apoptotic pathway is initiated by cytochrome c release from mitochondria. Deregulation of the cytochrome c apoptotic pathway can lead to diseases such as cancer, immune disorders, and neurodegenerative diseases. The overall goal of this project is to identify Apaf-1 inhibitors by high throughput screening. Apaf-1 is the essential mediato
- SAR analysis of small molecule activators of Apaf-1 using a LZ-Caspase-9/Caspase-3 Fluorescent Selectivity assay - Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBIMR, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: R01 CA136513 Assay Provider: Dr. Xuejun Jiang, Sloan-Kettering Institute for Cancer Research, New York, NY Apoptosis is a major form of programmed cell death that multicellular organisms utilize to maintain tissue homeostasis and to eliminate unwanted or damaged cells. It plays a critical role in development, immune responses and many other physiological events. In mammals, the mitochondrial cytochrome c-mediated apoptotic pathway is initiated by cytochrome c release from mitochondria. Deregulation of the cytochrome c apoptotic pathway can lead to diseases such as cancer, immune disorders, and neurodegenerative diseases. The overall goal of this project is to identify Apaf-1 activators by high throughput screening. Apaf-1 is the essential mediato
- SAR analysis of small molecule activators of the MazEF TA System via a fluorescence-based single-stranded RNase assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 2R01 GM068385-06 Assay Provider: Dr. Paul Hergenrother, University of Illinois, Urbana, IL Bacterial resistance to antibiotics is a worldwide health crisis. Resistance typically occurs as a result of chromosomal mutation or acquisition of a mobile genetic element, such as a plasmid, that harbors resistance-mediating genes. One of the most common plasmid maintenance systems is the toxin-antitoxin (TA) postsegregational killing mechanism. In this mechanism, if a plasmid-free daughter cell arises, the labile antitoxin is degraded and the toxin induces cell death. TA genes are ubiquitous in clinical isolates of certain drug-resistant bacteria, and it has been postulated that compounds that disrupt the TA interaction could free the toxin to kill the bac
- SAR analysis of small molecule agonists of the kappa opioid receptor via a luminescent beta-arrestin assay - Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak , Duke University, Durham NC Drug addiction is a disease originating in the central nervous system that produces compulsive behaviors despite the negative consequences that may result. Major addictive drugs of abuse include components of tobacco, opiates, marijuana, ethanol, cocaine, and derivatives of amphetamines. While the addictive behaviors produced by these substances may be generally similar, the drugs act at different receptor sites in the brain. Recent studies have shown that opioid receptors play a role regulating the addictive behaviors of other receptors that interact with illicit and legal substances of abuse. Opioid receptors are composed of multiple subtypes whose contributions to addicti
- SAR analysis of small molecule agonists of the kappa opioid receptor via a luminescent beta-arrestin assay - Set 3 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak , Duke University, Durham NC Drug addiction is a disease originating in the central nervous system that produces compulsive behaviors despite the negative consequences that may result. Major addictive drugs of abuse include components of tobacco, opiates, marijuana, ethanol, cocaine, and derivatives of amphetamines. While the addictive behaviors produced by these substances may be generally similar, the drugs act at different receptor sites in the brain. Recent studies have shown that opioid receptors play a role regulating the addictive behaviors of other receptors that interact with illicit and legal substances of abuse. Opioid receptors are composed of multiple subtypes whose contributions to addicti
- SAR analysis of small molecule agonists of the kappa opioid receptor via a luminescent beta-arrestin assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak , Duke University, Durham NC Drug addiction is a disease originating in the central nervous system that produces compulsive behaviors despite the negative consequences that may result. Major addictive drugs of abuse include components of tobacco, opiates, marijuana, ethanol, cocaine, and derivatives of amphetamines. While the addictive behaviors produced by these substances may be generally similar, the drugs act at different receptor sites in the brain. Recent studies have shown that opioid receptors play a role regulating the addictive behaviors of other receptors that interact with illicit and legal substances of abuse. Opioid receptors are composed of multiple subtypes whose contributions to addicti
- SAR analysis of small molecule antagonists of the kappa opioid receptor via a luminescent beta-arrestin assay - Set 3 Data Source: Burnham Center for Chemical Genomics (BCCG) Source Affiliation: Burnham Institute for Medical Research (BIMR, La Jolla, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak , Duke University, Durham NC Drug addiction is a disease originating in the central nervous system that produces compulsive behaviors despite the negative consequences that may result. Major addictive drugs of abuse include components of tobacco, opiates, marijuana, ethanol, cocaine, and derivatives of amphetamines. While the addictive behaviors produced by these substances may be generally similar, the drugs act at different receptor sites in the brain. Recent studies have shown that opioid receptors play a role regulating the addictive behaviors of other receptors that interact with illicit and legal substances of abuse. Opioid receptors are composed of multiple subtypes whose contributions to addictive behavio
- SAR analysis of small molecule antagonists of the kappa opioid receptor via a luminescent beta-arrestin assay - Set 4 Data Source: Burnham Center for Chemical Genomics (BCCG) Source Affiliation: Burnham Institute for Medical Research (BIMR, La Jolla, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak , Duke University, Durham NC Drug addiction is a disease originating in the central nervous system that produces compulsive behaviors despite the negative consequences that may result. Major addictive drugs of abuse include components of tobacco, opiates, marijuana, ethanol, cocaine, and derivatives of amphetamines. While the addictive behaviors produced by these substances may be generally similar, the drugs act at different receptor sites in the brain. Recent studies have shown that opioid receptors play a role regulating the addictive behaviors of other receptors that interact with illicit and legal substances of abuse. Opioid receptors are composed of multiple subtypes whose contributions to addictive behavio
- SAR analysis of small molecule antagonists of the kappa opioid receptor via a luminescent beta-arrestin assay - Set 5 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak , Duke University, Durham NC Drug addiction is a disease originating in the central nervous system that produces compulsive behaviors despite the negative consequences that may result. Major addictive drugs of abuse include components of tobacco, opiates, marijuana, ethanol, cocaine, and derivatives of amphetamines. While the addictive behaviors produced by these substances may be generally similar, the drugs act at different receptor sites in the brain. Recent studies have shown that opioid receptors play a role regulating the addictive behaviors of other receptors that interact with illicit and legal substances of abuse. Opioid receptors are composed of multiple subtypes whose contributions to ad
- SAR analysis of small molecule antagonists of the kappa opioid receptor via a luminescent beta-arrestin assay - Set 6 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network(MLPCN)Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak , Duke University, Durham NC Drug addiction is a disease originating in the central nervous system that produces compulsive behaviors despite the negative consequences that may result. Major addictive drugs of abuse include components of tobacco, opiates, marijuana, ethanol, cocaine, and derivatives of amphetamines. While the addictive behaviors produced by these substances may be generally similar, the drugs act at different receptor sites in the brain. Recent studies have shown that opioid receptors play a role regulating the addictive behaviors of other receptors that interact with illicit and legal substances of abuse. Opioid receptors are composed of multiple subtypes whose contributions to addi
- SAR analysis of small molecule antagonists of the kappa opioid receptor via a luminescent beta-arrestin assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak , Duke University, Durham NC Drug addiction is a disease originating in the central nervous system that produces compulsive behaviors despite the negative consequences that may result. Major addictive drugs of abuse include components of tobacco, opiates, marijuana, ethanol, cocaine, and derivatives of amphetamines. While the addictive behaviors produced by these substances may be generally similar, the drugs act at different receptor sites in the brain. Recent studies have shown that opioid receptors play a role regulating the addictive behaviors of other receptors that interact with illicit and legal substances of abuse. Opioid receptors are composed of multiple subtypes whose contributions to addictiv
- SAR analysis of small molecule inhibitors of Apaf-1 using a LZ-Caspase-9/Caspase-3 Fluorescent Selectivity assay - Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBIMR, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: R01 CA136513 Assay Provider: Dr. Xuejun Jiang, Sloan-Kettering Institute for Cancer Research, New York, NY Apoptosis is a major form of programmed cell death that multicellular organisms utilize to maintain tissue homeostasis and to eliminate unwanted or damaged cells. It plays a critical role in development, immune responses and many other physiological events. In mammals, the mitochondrial cytochrome c-mediated apoptotic pathway is initiated by cytochrome c release from mitochondria. Deregulation of the cytochrome c apoptotic pathway can lead to diseases such as cancer, immune disorders, and neurodegenerative diseases. The overall goal of this project is to identify Apaf-1 inhibitors by high throughput screening. Apaf-1 is the essential mediato
- SAR analysis of small molecule inhibitors of tim23-1 yeast via a luminescent assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 DA027714-01A1 Assay Provider: Dr. Carla Koehler, University of California, Los Angeles, CA (UCLA) Defects in mitochondrial assembly impact a wide range of diseases from degenerative muscle and neural diseases to cancer (Wallace, 2005). The mitochondrion is not only important for the production of energy but plays an important role in other aspects such as intermediary metabolism and signaling. The mitochondrion contains an inner membrane and outer membrane that separate the matrix from the intermembrane space. Proteins destined for the mitochondrion are imported via the Translocase of the Outer Membrane (TOM) and the Translocases of the Inner Membrane (TIM23 for proteins destined for the matrix and TIM22 for proteins destined for the inner me
- SAR analysis of small molecule inhibitors of tim23-1: a luminescent TIM10 yeast counterscreen. Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 DA027714-01A1 Assay Provider: Dr. Carla Koehler, University of California, Los Angeles, CA (UCLA) Defects in mitochondrial assembly impact a wide range of diseases from degenerative muscle and neural diseases to cancer (Wallace, 2005). The mitochondrion is not only important for the production of energy but plays an important role in other aspects such as intermediary metabolism and signaling. The mitochondrion contains an inner membrane and outer membrane that separate the matrix from the intermembrane space. Proteins destined for the mitochondrion are imported via the Translocase of the Outer Membrane (TOM) and the Translocases of the Inner Membrane (TIM23 for proteins destined for the matrix and TIM22 for proteins destined for the inner me
- SAR analysis of small molecule inhibitors of tim23-1: a luminescent tim10-1 yeast counterscreen. Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 DA027714-01A1 Assay Provider: Dr. Carla Koehler, University of California, Los Angeles, CA (UCLA) Defects in mitochondrial assembly impact a wide range of diseases from degenerative muscle and neural diseases to cancer (Wallace, 2005). The mitochondrion is not only important for the production of energy but plays an important role in other aspects such as intermediary metabolism and signaling. The mitochondrion contains an inner membrane and outer membrane that separate the matrix from the intermembrane space. Proteins destined for the mitochondrion are imported via the Translocase of the Outer Membrane (TOM) and the Translocases of the Inner Membrane (TIM23 for proteins destined for the matrix and TIM22 for proteins destined for the inner me
- SAR assay for compounds activating TNAP in the absence of phosphate acceptor performed in a luminescent assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Proposal Number: 1R03 MH082385-01 This TNAP dose response assay is developed and performed to confirm hits originally identified in the TNAP luminescent HTS assay (AID 1001) and to study the structure-activity relationship on analogs of the confirmed hits. Compounds are either acquired from commercial sources or synthesized internally. Alkaline phosphatases (EC 3.1.3.1) (APs) catalyze the hydrolysis of phosphomonoesters, releasing phosphate and alcohol. APs are dimeric enzymes found in the most organisms. In human, four isozymes of APs have been identified. Three isozymes are tissue-specific and the fourth one is tissue-non specific, named TNAP. TNAP deficiency is associated with defective bone mineralization in the form of rickets and osteomalacia. Therefore, t
- SAR assay for compounds activating TNAP in the presence of 100 mM DEA performed in a luminescence assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Proposal Number: 1R03 MH082385-01 This TNAP dose response assay is developed and performed to confirm hits originally identified in the TNAP luminescent HTS assay (AID 1001) and to study the structure-activity relationship on analogs of the confirmed hits. Compounds are either acquired from commercial sources or synthesized internally. Alkaline phosphatases (EC 3.1.3.1) (APs) catalyze the hydrolysis of phosphomonoesters, releasing phosphate and alcohol. APs are dimeric enzymes found in the most organisms. In human, four isozymes of APs have been identified. Three isozymes are tissue-specific and the fourth one is tissue-non specific, named TNAP. TNAP deficiency is associated with defective bone mineralization in the form of rickets and osteomalacia. Therefore, t
- TR-FRET secondary assay for HTS discovery of chemical inhibitors of Hsp70 Sanford-Burnham Center for Chemical Genomics (SBCCG) Sanford-Burnham Medical Research Institute (San Diego, CA) NIH Molecular Libraries Screening Centers Network (MLSCN) MLSCN Grant: XO1 MH079863-01 Over-expression of molecular chaperones occurs commonly in cancers and provides protection from a wide variety of cellular stresses, both endogenous and iatrogenic. Molecular chaperones also play important roles in maintaining the activity of several signal-transducing proteins and transcriptions factors involved in malignant transformation. The human genome contains nine Hsp70-family genes. These chaperones include Hsp70 and Hsc70, which are commonly over-expressed in cancers and which confer resistance to myriad cellular stresses, including cytotoxic chemotherapy. The current assay was developed at the Sanford-Burnham Center for Chemical Genomics (SBCCG), based on Fluorescen-12-ATP binding to GST-Hsp70 in the presence of Terbium-labeled anti-GST antibody. The assay is aimed to support
- uHTS HTRF assay for identification of inhibitors of SUMOylation Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH084862-01 Assay Provider: Dr. Yuan Chen, Beckman Research Institute, City Of Hope, CA Protein modification by the SUMO (Small Ubiquitin-like MOdifier) family of proteins is an important post-translational modification that plays an essential role in many functions including gene transcription, cell cycle progression, DNA repair, viral infection, and the development of neurodegenerative diseases (1, 2). Recent proteomic studies have found that approximately 10% of the proteins encoded by the yeast genome are substrates for SUMO modification (3-5). The mechanism of how SUMOylation is involved in these cellular functions remains largely unclear. The inhibitors of SUMOylation would be useful to probe the roles of SUMOylation in cellular regulati
- uHTS identification of compounds activating TNAP in the absence of phosphate acceptor performed in luminescent assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Proposal Number: 1R03 MH082385-01 Alkaline phosphatases (EC 3.1.3.1) (APs) catalyze the hydrolysis of phosphomonoesters, releasing phosphate and alcohol. APs are dimeric enzymes found in the most organisms. In human, four isozymes of APs have been identified. Three isozymes are tissue-specific and the fourth one is tissue-non specific, named TNAP. TNAP deficiency is associated with defective bone mineralization in the form of rickets and osteomalacia. Therefore, there is therapeutic potential of modulating TNAP activity. The goal of this HTS is to identify novel and specific activators of TNAP. The only known to date class of alkaline phosphatases activators are amino-containing alcohols, such as diethanolamine (DEA), that act as phosphoacceptor substrate and exh
- uHTS identification of small molecule antagonists of the kappa opioid receptor via a luminescent beta-arrestin assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1X01DA026208-01 Assay Provider: Dr. Lawrance Barak , Duke University, Durham NC Drug addiction is a disease originating in the central nervous system that produces compulsive behaviors despite the negative consequences that may result. Major addictive drugs of abuse include components of tobacco, opiates, marijuana, ethanol, cocaine, and derivatives of amphetamines. While the addictive behaviors produced by these substances may be generally similar, the drugs act at different receptor sites in the brain. Recent studies have shown that opioid receptors play a role regulating the addictive behaviors of other receptors that interact with illicit and legal substances of abuse. Opioid receptors are composed of multiple subtypes whose contributions to addictiv
- uHTS identification of small molecule inhibitors of LYP via a fluorescence intensity assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1R21NS056945-01 Assay Provider: Dr. Nunzio Bottini, San Diego Institute for Allergy and Immunology CA LYP, is a lymphocyte specific protein tyrosine phosphatase that plays a critical regulatory role in T cell receptor signaling. The PTPN22 gene encodes this phosphatase. A single-nucleotide polymorphism in PTPN22 is associated with a number of autoimmune disorders, including type 1 diabetes, rheumatoid arthritis, juvenile rheumatoid arthritis, systemic lupus erythematosus and Grave's disease. The autoimmunity-predisposing allele is a gain-of-function mutant suggesting that a specific small-molecule inhibitor could eliminate its effect. Finding specific inhibitors of protein phosphatases has proven extremely difficult. The goal of this project is to f
- Dose Response confirmation of APOBEC3A DNA Deaminase Inhibitors via a A3G counterscreen Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH089432-01 Assay Provider: Dr. Reuben S Harris, Regents of the University of Minnesota, Minneapolis, MN The human APOBEC3G (A3G) protein is a DNA deaminase that has potent activity against HIV-1 and a variety of other retroelements. Although HIV encodes a natural inhibitor of A3G called Vif, it was hypothesized that the virus still benefits from the mutagenic activity of A3G (immune escape and drug resistance). The purpose of the screening is to identify A3G inhibitor by using a fluorescence-based single-strand DNA deaminase assay. In this assay, an A3G protein, a single-strand DNA oligonucleotide with a 5'-CCC target site, and Uracil DNA Glycosylase (UDG) are all mixed. After incubation at room temperature, A3G deaminates C-to-U
- Dose Response confirmation of APOBEC3G DNA Deaminase Inhibitors via a A3A counterscreen Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH089432-01 Assay Provider: Dr. Reuben S Harris, Regents of the University of Minnesota, Minneapolis, MN The human APOBEC3G (A3G) protein is a DNA deaminase that has potent activity against HIV-1 and a variety of other retroelements. Although HIV encodes a natural inhibitor of A3G called Vif, it was hypothesized that the virus still benefits from the mutagenic activity of A3G (immune escape and drug resistance). The purpose of the screening is to identify A3G inhibitor by using a fluorescence-based single-strand DNA deaminase assay. In this assay, an A3G protein, a single-strand DNA oligonucleotide with a 5'-CCC target site, and Uracil DNA Glycosylase (UDG) are all mixed. After incubation at room temperature, A3G deaminates C-to-U
- Dose Response confirmation of Inhibitors of Mdm2/MdmX interaction using a Brca1/Bard1 BiLC Counterscreen assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: R03 MH089489-01 Assay Provider: Dr. Geoffrey M. Wahl, Salk Institute for Biological Studies, San Diego, CA A wild type but attenuated p53 is retained in approximately 50% of human tumors, and reactivation of p53 in such tumors is an attractive chemotherapeutic strategy. p53 activity is restricted in vivo by mdm2 and mdmx, and knockout of either of these proteins is embryonic lethal in a p53-dependent manner (1, 2). Both proteins bind to p53 via a hydrophobic N-terminal pocket and block p53-dependent transcription of genes required for tumor suppression. Efforts to reactivate p53 with small molecules have focused on inhibition of the mdm2/p53 interaction, which leads to increased p53 levels and activity. However, recent reports indicate that targetin
- Dose Response confirmation of small molecule APOBEC3A DNA Deaminase Inhibitors via a fluorescence-based single-stranded DNA deaminase assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH089432-01 Assay Provider: Dr. Reuben S Harris, Regents of the University of Minnesota, Minneapolis, MN The human APOBEC3G (A3G) protein is a DNA deaminase that has potent activity against HIV-1 and a variety of other retroelements. Although HIV encodes a natural inhibitor of A3G called Vif, it was hypothesized that the virus still benefits from the mutagenic activity of A3G (immune escape and drug resistance). The purpose of the screening is to identify A3G inhibitor by using a fluorescence-based single-strand DNA deaminase assay. In this assay, an A3G protein, a single-strand DNA oligonucleotide with a 5'-CCC target site, and Uracil DNA Glycosylase (UDG) are all mixed. After incubation at room temperature, A3G deaminates C-to-U an
- Dose Response confirmation of small molecule APOBEC3G DNA Deaminase Inhibitors via a fluorescence-based single-stranded DNA deaminase assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH089432-01 Assay Provider: Dr. Reuben S Harris, Regents of the University of Minnesota, Minneapolis, MN The human APOBEC3G (A3G) protein is a DNA deaminase that has potent activity against HIV-1 and a variety of other retroelements. Although HIV encodes a natural inhibitor of A3G called Vif, it was hypothesized that the virus still benefits from the mutagenic activity of A3G (immune escape and drug resistance). The purpose of the screening is to identify A3G inhibitor by using a fluorescence-based single-strand DNA deaminase assay. In this assay, an A3G protein, a single-strand DNA oligonucleotide with a 5'-CCC target site, and Uracil DNA Glycosylase (UDG) are all mixed. After incubation at room temperature, A3G deaminates C-to-U
- Dose Response confirmation of small molecule inhibitors originally identified via uHTS of Artemis endonuclease activity via a fluorescence intensity assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH095489-01A1 Assay Provider: Michael Lieber, M.D., Ph.D., University of Southern California, Los Angeles, CA Nonhomologous DNA end joining (NHEJ) is the primary DNA repair pathway in human cells for the repair of double-strand DNA breaks. Like most DNA repair pathways, NHEJ relies on three enzyme activities: a nuclease to remove damaged DNA, polymerases to fill-in new DNA, and a ligase to restore DNA strand integrity. The primary nuclease for NHEJ is Artemis, which is activated when a protein kinase called DNA-PKcs makes contact with a double-stranded DNA end at a chromosomal break site. Artemis is an endonuclease which nicks the DNA at transitions between single- and double-stranded DNA. It is very important for repairing a critical subset o
- Dose response confirmation uHTS hits for MazEF TA System activators via a fluorescence-based single-stranded RNase assay counterscreen Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 2R01 GM068385-06 Assay Provider: Dr. Paul Hergenrother, University of Illinois, Urbana, IL The assay was developed and performed in dose-response as a counter assay for the primary assay originally identified as "identification of MazEF TA System activators via a fluorescence-based single-stranded RNase assay", AID 504720. The goal of the assay is to identify compounds that exhibit an increase in fluorescence over the timecourse of the assay in the absence of the MazEF TA system. The counter dose-response assay described here will enable us to filter fluorescent compounds and readily identify small molecules that activate RNase-based toxins. A fluorophore and a quencher are on the opposite ends of an oligonucleotide substrate. When the RNase (i
- Dose-response secondary confirmation of microRNA-mediated mRNA deadenylation inhibitors by fluoresence polarization assay using Cy5 labeled peptide Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network(MLPCN) Grant Number: R03 MH094198-01 Assay Provider: Kalle Gehring, Ph.D., McGill University The poly(A) tail of mRNA is bound by several molecules of the poly(A)-binding protein (PABP), an abundant cytoplasmic protein in eukaryotes that promotes translation (2). PABPC1 is a multi-domain protein that contains four phylogenetically conserved RNA recognition motifs (RRM1-4) (3,4) a proline-rich unstructured region and a 70-residue C-terminal domain termed Mlle or PABC (5). Mlle (Mademoiselle) refers to the conserved KITGMLLE signature motif of this domain. The Mlle domain binds proteins with a 12-15 amino acid motif termed PAM2 to regulate PABPC1 involvement in mRNA decay. The goal of this grant is to screen for small molecule modulators of Mlle interactions as chemical probes
- MKP-3 in vitro secondary assay for identification of irreversible and slow-binding inhibitors Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute(SBMRI, San Diego, CA) Network: NIH Molecular Libraries Screening Centers Network (MLSCN) Grant Number: XO1 MH076390-01 Assay Provider: Dr. John Lazo, University of Pittsburg This assay is designed as a counter-screen for the MKP-3 in vitro HTS assay (AID 425) aimed at identification of compounds with time-dependent behavior. MKP-3 (mitogen-activated protein kinase phosphatase-3; EC 3.1.3.48, EC 3.1.3.16), a dual specificity phosphatase negatively regulates ERK1/2 by catalyzing the removal of a phosphoryl group from Thr(P) and Tyr(P) in the activation loop consensus motif -pTXpY. MKP-3 has a labile cysteine in its active site that is crucial for the catalysis. As a result compounds capable of cysteine oxidation or modification are likely to appear as screening hits. The current biochemical assay is designed as a rapid diagnostic tool for identification of
- SAR analysis of compounds that inhibit Human Immunodeficiency Virus Fusion, cell-cell fusion assay Data Source: Burnham Center for Chemical Genomics (BCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Production Centers Network (MLPCN) Grant Number: 1R21NS059403-01 Assay Provider: Dr. Miriam Gochin, Touro University-California, Vallejo, CA The fusion-active conformation of the envelope protein gp41 of HIV-1 consists of an N-terminal trimeric a-helical coiled coil domain, and three anti-parallel C-terminal helices which fold down the grooves of the coiled coil to form a six-helix bundle. Disruption of the six-helix bundle is considered to be a key component of an effective non-peptide fusion inhibitor. This structure forms as a result of a conformational change in gp41, triggered by gp120 and co-receptor binding to host cell receptors. Prevention of six-helix bundle formation has been recognized as an important mechanism for viral fusion inhibition This confirmatory, concentration-response assay has been de
- SAR analysis of small molecule inhibitors of APOBEC3A DNA Deaminase via a fluorescence-based single-stranded DNA deaminase assay - Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH089432-01 Assay Provider: Dr. Reuben S Harris, Regents of the University of Minnesota, Minneapolis, MN The human APOBEC3G (A3G) protein is a DNA deaminase that has potent activity against HIV-1 and a variety of other retroelements. Although HIV encodes a natural inhibitor of A3G called Vif, it was hypothesized that the virus still benefits from the mutagenic activity of A3G (immune escape and drug resistance). The purpose of the screening is to identify A3G inhibitor by using a fluorescence-based single-strand DNA deaminase assay. In this assay, an A3A protein, a single-strand DNA oligonucleotide with a 5'-CCC target site, and Uracil DNA Glycosylase (UDG) are all mixed. After incubation at room temperature, A3G deaminates C-to-U and
- SAR analysis of small molecule inhibitors of APOBEC3G DNA Deaminase via a fluorescence-based single-stranded DNA deaminase assay - Set 2 Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH089432-01 Assay Provider: Dr. Reuben S Harris, Regents of the University of Minnesota, Minneapolis, MN The human APOBEC3G (A3G) protein is a DNA deaminase that has potent activity against HIV-1 and a variety of other retroelements. Although HIV encodes a natural inhibitor of A3G called Vif, it was hypothesized that the virus still benefits from the mutagenic activity of A3G (immune escape and drug resistance). The purpose of the screening is to identify A3G inhibitor by using a fluorescence-based single-strand DNA deaminase assay. In this assay, an A3G protein, a single-strand DNA oligonucleotide with a 5'-CCC target site, and Uracil DNA Glycosylase (UDG) are all mixed. After incubation at room temperature, A3G deaminates C-to-U and
- SAR analysis of small molecule inhibitors of APOBEC3G DNA Deaminase via a fluorescence-based single-stranded DNA deaminase assay Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG) Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA) Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN) Grant Number: 1 R03 MH089432-01 Assay Provider: Dr. Reuben S Harris, Regents of the University of Minnesota, Minneapolis, MN The human APOBEC3G (A3G) protein is a DNA deaminase that has potent activity against HIV-1 and a variety of other retroelements. Although HIV encodes a natural inhibitor of A3G called Vif, it was hypothesized that the virus still benefits from the mutagenic activity of A3G (immune escape and drug resistance). The purpose of the screening is to identify A3G inhibitor by using a fluorescence-based single-strand DNA deaminase assay. In this assay, an A3G protein, a single-strand DNA oligonucleotide with a 5'-CCC target site, and Uracil DNA Glycosylase (UDG) are all mixed. After incubation at room temperature, A3G deaminates C-to-U and
- TR-FRET secondary assay for HTS discovery of chemical inhibitors of anti-apoptotic protein Bfl-1 Sanford-Burnham Center for Chemical Genomics (SBCCG) Sanford-Burnham Medical Research Institute (San Diego, CA) NIH Molecular Libraries Screening Centers Network (MLSCN) Bfl-1, also known as A1 in mice is an anti-apoptotic and NF-kB-inducible member of the Bcl-2 protein family involved in regulation of apoptosis. Due to difficulties with accomplishing targeted gene ablation in mouse models, the endogenous functions of Bfl-1 are largely unknown. Chemical inhibitors of Bfl-1 can be used as research tools for neutralizing Bfl-1 in human and mouse cells. The current assay was developed at the Sanford-Burnham Center for Chemical Genomics (SBCCG), based on FITC-Bid BH3 peptide binding to GST-Bfl-1 in the presence of Terbium-labeled anti-GST antibody. The assay is aimed to support Bfl-1 chemical probe identification through the TR-FRET assay confirmation of the results obtained in the primary fluorescence polarization (FP) assay (PubChem AID 432) performed at the BCCG. Bfl-1 screening w
- Enzyme Activity Inhibition Assay The scintillation Proximity Assay (SPA) method was used to determine the inhibitory effect of compounds on the activity of PDE4D catalytic domain. Human PDE4D catalytic domain protein was obtained by expression and purification in E. coli. The positive compound Apremilast was purchased from Topscience Biochemical, Microplate Scintillation Counter (MicroBeta2, Perkin Elmer), constant temperature water bath (DK420, Shanghai Medical Device Factory), Micro-vibrator (XW-80A, Shanghai Jingke Industrial Co., Ltd.) are a public instrument in the radioactive laboratory, and the step-by-step pipette (Multipette Plus, Eppendorf) and supporting tips were purchased from Ebende Biotechnology Company, 3.5. [3H]-cAMP, scintillation beads (RPNQO150, Perkin Elmer), and 96-well scintillation microplates (Isoplate-96, Perkin Elmer) were purchased from Perkin Elmer Company. 10× SPA buffer was prepared in the laboratory (500 mM Tris pH7.5, 83 mM MgCl2, 17 mM EGTA).In the experiment, 60 μl water and 10 μl reaction solution were added into 100 μl total volume reaction to achieve the final concentration of each component being 50 mM Tris-HCl, pH7.5, 8.3 mM MgCl2, 1.7 mM EGTA, 10 μl compound and 10 μl enzyme (0.1 ng/ul). Finally, 10 μl [3H]-cAMP (0.005 μCi/μl) was added and incubated for 30 min at 30° C. in a water bath. 50 μl SPA beads was added to quench the reaction, shaked appropriately, and stood for 20 minutes. A microplate scintillation counter was used
- Inhibitory Activity against 3CLPpro The inhibitory activity of the compounds against SARS-CoV-2 3CLpro was determined by fluorescence resonance energy transfer technique. A suitable amount of the above compounds was respectively weighed, and formulated in DMSO to give solutions over suitable concentration gradients. 5 μL of the formulated solution and 91 μL of Assay Reagent (Assay Buffer: 2019-nCoV Mpro/3CLpro=90:1, purchased from Shanghai Beyotime Biotechnology Co., Ltd.) were added to a 96-well black plate, mixed uniformly, and incubated for 10 min at 37° C. in the dark. 4 μL of Substrate (100 μM Dabcyl-KTSAVLQSGFRKME-Edans, purchased from Shanghai Beyotime Biotechnology Co., Ltd.) was rapidly added to each well, and mixed uniformly. After incubation for 5 min at 37° C. in the dark, the signal gradually became stable. The fluorescence in 5-30 min was detected on a multifunctional plate reader (Thermo Fisher Technology Co., LTD., Varioskan Flash) and the inhibition percentage of the sample was calculated. The excitation wavelength was 340 nm and the emission wavelength was 490 nm. The Assay Reagent free of the compound was used as a control with 100% enzyme activity, the Assay Buffer free of SARS-CoV-2 Mpro/3CLpro was used as a blank control, and S-216722 (Shandong Xuanshuo Medical Technology Co., Ltd.) and PF-07321332 (Jinan Jianfeng Chemical Co., Ltd.) were used as positive controls. The rest of the treatment method was the same.
- Biochemical Assay 384-well (Greiner #784075) (or 1536-well [Greiner #782075]) plates containing 60 nl (or 20 nl for 1536-well) of compound in 100% DMSO (10 concentrations serially diluted 3.16-fold) were prepared. Two copies of plates were used one for the GCN2 assay and a second for an artifact plate. Working reagents were prepared as follows reaction buffer: Tris pH 7.5 20 mM, MgCl2 5 mM, DTT 1 mM, Brij-35 0.005%, EGTA 0.5 mM in water; 2 enzyme solution: GCN2 (produced in-house) 1.6 nM in reaction buffer; 2 substrate solution: GFP-eIFS1 (Thermo Fisher Scientific, PV4809) 50 nM, ATP 0.2 mM, tRNA 0.4 mg/ml in reaction buffer; p-GFP-eIF2S1 (artifact) solution: p-GFP-eIFS1 (produced by incubation of 100 nM GFP-eIF2S1 in reaction buffer with 100 uM ATP and 1 nM PERK KD enzyme for 3 hr at room temperature) 2 nM in reaction buffer; 3 stop/antibody solution: p-GFP-EIF2S1 Th-Ab (Thermo Fisher Scientific, PV4816) 6 nM, AZ13933939 3 uM, BSA 3% in reaction buffer. Reagents were loaded onto a liquid dispenser (Certus FLEX, Trajan Scientific and Medical), and 3 ul (1 ul for 1536-well) of 2 enzyme solution, followed by 3 ul (1 ul for 1536-well) of 2 substrate solution was added to the plates. When small numbers of plates were processed, the enzyme solution was added first to all plates, then the valve used for the substrate solution was primed immediately prior to use. Plates were tapped or spun 1 min/1K/RT to ensure proper mixing, followed by incubation at room temperature for 70 mins, covered/stacked in the dark.
- Beta-2 AR binding assay HEK 293 cells stability transfected with cDNA encoding human β2-AR (provided by Dr. Brian Kobilka, Stanford Medical Center, Palo Alto, Calif.) were grown in Dulbecco's Modified Eagle Medium (DMEM) containing 10% fetal bovine serum (FBS), 0.05% penicillin-streptomycin, and 400 μg/ml G418 as previously described (Pauwels et al., Biochem. Pharmacol. 42: 1683-1689, 1991). The cells were scraped from the 150×25 mm plates and centrifuged at 500×g for 5 minutes. The pellet was homogenized in 50 mM Tris-HCl, pH 7.7, with a Polytron, centrifuged at 27,000×g, and resuspended in the same buffer. The latter process was repeated, and the pellet was resuspended in 25 mM Tris-HCl containing 120 mM NaCl, 5.4 mM KCl, 1.8 mM CaCl2, 0.8 mM MgCl2, and 5 mM glucose, pH 7.4. The binding assays contained 0.3 nM [3H]CGP-12177 in a volume of 1.0 ml. Nonspecific binding was determined by 1 μM propranolol.According to the above-described methods, binding affinities, expressed as Ki values, were determined using membranes obtained from a HEK 293 cell line stably transfected with cDNA encoding human β2-AR (Pauwels et al., Biochem. Pharmacol. 42: 1683-1689, 1991) with [3H]CGP-12177 as the marker ligand. The resulting IC50 values and Hill coefficients were calculated for each test compound using GraphPad Prism software and Ki values were calculated using the Cheng-Prusoff transformation (Biochem Pharmacol 22: 3099-3108, 1973):K i=IC50/(1+L/K d)+ Eqn. 1.Where: L is the concentration of [3H]CGP-12177 and Kd is the binding affinity of the [3H]CGP-12177. Each test compounds was assayed three times.
- In Vitro Detection of the Inhibitory Activity of Compounds Against PDE 3A Enzyme Experimental objective: to determine the AMP/GMP expression based on fluorescence polarization, i.e., to trace binding of AMP/GMP to antibody so as to indicate enzyme activity.Reagents:Experimental buffer solution: 10 mM Tris-HCl (pH 7.5), 5 mM MgCl2, 0.01% Brij 35, 1 mM Dithiothreitol (DTT), and 1% DMSO.Enzyme: recombinant human PDE3A (Gene accession number: NM_000921; amino acid 669-end) was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag, with molecular weight being 84 kDa.Enzyme substrate: 1 μM cAMPDetection: Transcreener AMP2/GMP2 antibody and AMP2/GMP2 AlexaFluor633 tracer.Procedure:1. The recombinant human PDE3A enzyme and enzyme substrate (1 μM cAMP) were each dissolved in newly-prepared experimental buffer solution;2. The PDE3A enzyme buffer solution was transferred into reaction wells;3. The compound which was dissolved in 100% DMSO was added to the reaction wells containing PDE3A enzyme buffer solution by acoustic technique (echo 550; millilambda range) and the mixture was incubated for 10 minutes at room temperature;4. The enzyme substrate buffer solution was added to the above reaction wells to initiate reaction;5. The resulting mixture was incubated at room temperature for 1 hour;6. The detection mixture (Transcreener AMP2/GMP2 antibody and AMP2/GMP2 AlexaFluor633 tracer) was added to stop the reaction, and the resulting mixture was incubated for 90 minutes while slowly mixing. The measurement range of fluorescence polarization was Ex/Em=620/688.Data analysis: the fluorescence polarization signal was converted to nM based on AMP/GMP standard curve and the percentage enzyme activity relative to DMSO control calculated by Excel. GraphPad Prism was used for curve fitting (drawing medical icon).
- Inhibitory Activity Against IDH1R132H and IDH1R132C Enzymes The IDH1R132H protein and the IDH1R132C protein were prepared as follows: the IDH1R132H or IDH1R132C gene was integrated into a pET24b(+) vector (Novagen) to prepare a construct for C-terminal fusion of 6× histidine tag. After transformation of Rosetta 2 (DE3) E. coli, the expression of the protein was induced with IPTG. The E. coli was collected and homogenized, followed by the affinity purification of the 6× histidine fusion protein using HisTrap HP columns (GE Healthcare Japan Corp.) and gel filtration purification using Superdex 200 columns (GE Healthcare Japan Corp.) to obtain the IDH1R132H or IDH1R132C protein of interest.IDH1R132H and IDH1R132C each convert 2-oxoglutarate and NADPH to D-2-hydroxyglutarate (2-HG) and NADP+. Therefore, the activity of the IDH1R132H and IDH1R132C enzymes can be measured by detecting NADPH levels.The enzyme inhibitory activity evaluation was carried out as follows: 40 μL each of reaction solutions containing different concentrations of each compound (100 mM Tris-HCl (pH 7.5), 150 mM NaCl, 20 mM MgCl2, 0.5 mg/mL bovine serum albumin, 1 mM reduced glutathione, 40 μM NADPH, 0.5 mM 2-oxoglutarate, 0.5% dimethyl sulfoxide, 50000 to 0.128 nM compound, and 12 nM IDH1R132H or 10 nM IDH1R132C as the enzyme) was added to each well of 384-well plates (Greiner Bio One International GmbH, #781096) and incubated at room temperature. While NADPH-derived fluorescence was occasionally monitored, the reaction was terminated by the addition of 5 μL of 0.5 M EDTA before consumption of NADPH. 5 μL of WST-8 reagent (Dojindo Laboratories, #CK04) was further added and mixed therewith. 15 minutes later, the absorbance at 450 nm was measured using a plate reader (PerkinElmer, Inc., EnVision). The observed absorbance value reflects the amount of residual NADPH. From the absorbance data, the enzyme inhibitory activity of each compound at each concentration was calculated and analyzed using medical statistical analysis software GraphPad Prism to calculate an IC50 value.