Development of a high-throughput screening method for LIM kinase 1 using a luciferase-based assay of ATP consumption.

ABSTRACT: Kinases are attractive drug targets because of the central roles they play in signal transduction pathways and human diseases. Their well-formed adenosine triphosphate (ATP)-binding pockets make ideal targets for small-molecule inhibitors. For drug discovery purposes, many peptide-based kinase assays have been developed that measure substrate phosphorylation using fluorescence-based readouts. However, for some kinases these assays may not be appropriate. In the case of the LIM kinases (LIMK), an inability to phosphorylate peptide substrates resulted in previous high-throughput screens (HTS) using radioactive labeling of recombinant cofilin protein as the readout. We describe the development of an HTS-compatible assay that measures relative ATP levels using luciferase-generated luminescence

SUBMITTER: Mezna M 

PROVIDER: S-EPMC3935456 | biostudies-literature | 2012 Apr

REPOSITORIES: biostudies-literature