High-throughput cell-based screening using scintillation proximity assay for the discovery of inositol phosphatase inhibitors

J Biomol Screen. 2004 Mar;9(2):132-40. doi: 10.1177/1087057103261039.

Abstract

Inositol monophosphatase is a potential drug target for developing lithium-mimetic agents for the treatment of bipolar disorder. Enzyme-based assays have been traditionally used in compound screening to identify inositol monophosphatase inhibitors. A cell-based screening assay in which the compound needs to cross the cell membrane before reaching the target enzyme offers a new approach for discovering novel structure leads of the inositol monophosphatase inhibitor. The authors have recently reported a high-throughput measurement of G-protein-coupled receptor activation by determining inositol phosphates in cell extracts using scintillation proximity assay. This cell-based assay has been modified to allow the determination of inositol monophosphatase activity instead of G-protein-coupled receptors. The enzyme is also assayed in its native form and physiological environment. The authors have applied this cell-based assay to the high-throughput screening of a large compound collection and identified several novel inositol monophosphatase inhibitors.

MeSH terms

  • Atropine / pharmacology
  • Biological Assay / methods
  • Carbachol / pharmacology
  • Cells, Cultured
  • Chromatography, High Pressure Liquid
  • Humans
  • Models, Biological
  • Phosphoric Monoester Hydrolases / antagonists & inhibitors*
  • Scintillation Counting

Substances

  • Atropine
  • Carbachol
  • Phosphoric Monoester Hydrolases
  • myo-inositol-1 (or 4)-monophosphatase