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Cell Reactant:
Major Urinary Protein (MUP-1)
Syringe Reactant:
BDBM22602
Meas. Tech.:
Isothermal Titration Calorimetry
Entry Date.:
06/14/08
ΔG°:
-17.2±0.3 (kJ/mole)
pH:
7.4±n/a
Log10Kb:
2.5
Temperature:
300±n/a (K)
ΔHobs :
-34.3±0.3 (kJ/mole)
Corrected for ΔHioniz:
no
Stoich. Param.:
0.95
ΔS° :
-0.057±0.0013 (kJ/mole-K)
Comments:
The affinity of 1,8 octan-diol for MUP-I was too weak to enable an accurate determination of thermodynamic binding parameters by direct titration, and hence the displacement ITC approach was used with 2-methoxy-3-isobutylpyrazine (IBMP) as the tight binding ligand. In a typical experiment, IBMP was in the injection syringe, and titrated into the mixture of MUP-1 and 1,8 octan-diol in the cell.
Citation
Humber, DC; Barratt, E; Bamford, MJ; Bronowska, A; Vondrásek, J; Bethell, RC; Cammack, N; Cerný, J; Bingham, R; Cobley, K; Evans, DN; Phillips, S; Gray, NM; Homans, SW; Hann, MM; Orr, DC; Saunders, J Thermodynamic penalty arising from burial of a ligand polar group within a hydrophobic pocket of a protein receptor. J Mol Biol 362:994-1003 (2006) [PubMed] Article
Cell React
Source:
The MUP-1 gene was cloned and expressed in E. coli cells.
Prep. Method:
MUP-I was purified using Ni-NTA resin. To remove endogenous bound ligands, an ethanol precipitation step was utilized involving addition of 2 volumes of absolute ethanol per volume of protein solution, followed by incubation at 4 °C for 2 h. After centrifugation at 5000g, the pellet was lyophilised, resuspended in PBS pH 7.4, and dialyzed overnight against the same buffer.
Name:
Major Urinary Protein (MUP-1)
Synonyms:
MUP1_MOUSE | Major urinary protein 1 precursor | Mup1
Type:
Other Protein Type
Mol. Mass.:
20640.17
Organism:
Mus musculus (mouse)
Description:
n/a
Residue:
180
Sequence:
MKMLLLLCLGLTLVCVHAEEASSTGRNFNVEKINGEWHTIILASDKREKIEDNGNFRLFLEQIHVLENSLVLKFHTVRDEECSELSMVADKTEKAGEYSVTYDGFNTFTIPKTDYDNFLMAHLINEKDGETFQLMGLYGREPDLSSDIKERFAQLCEKHGILRENIIDLSNANRCLQARE