Cracking the molecular weight barrier: fragment screening of an aminotransferase using an NMR-based functional assay

Bioorg Med Chem Lett. 2011 Sep 15;21(18):5248-50. doi: 10.1016/j.bmcl.2011.07.039. Epub 2011 Jul 21.

Abstract

NMR-based screening of protein targets has become a well established part of the drug discovery process especially with respect to fragments. However, as target size increases the two-dimensional spectra typically used for such screening become more crowded due to the increased number of signals, and the individual signals broaden due to the decreased rotational correlation time of the protein. Here we present an NMR-based functional assay for the branched-chain aminotransferase BCATc, a dimer with a total molecular weight of 88 kDa, which overcomes the limitations of the typical protein-based NMR screening method. BCATc is involved in glutamate production in the brain and is a therapeutic target for neuronal disorders involving a glutamatergic mechanism. Several fragments which inhibit BCATc were discovered using this assay and these may serve as novel cores for the development of potent BCATc inhibitors.

MeSH terms

  • Biocatalysis
  • Carbon Isotopes
  • Dose-Response Relationship, Drug
  • Enzyme Inhibitors / chemistry
  • Enzyme Inhibitors / pharmacology*
  • Ligands
  • Molecular Structure
  • Molecular Weight
  • Nuclear Magnetic Resonance, Biomolecular / methods*
  • Organic Chemicals / chemistry
  • Organic Chemicals / pharmacology*
  • Structure-Activity Relationship
  • Transaminases / antagonists & inhibitors*
  • Transaminases / chemistry
  • Transaminases / metabolism

Substances

  • Carbon Isotopes
  • Enzyme Inhibitors
  • Ligands
  • Organic Chemicals
  • Transaminases