Bisubstrate fluorescent probes and biosensors in binding assays for HTS of protein kinase inhibitors

Biochim Biophys Acta. 2010 Mar;1804(3):541-6. doi: 10.1016/j.bbapap.2009.10.019. Epub 2009 Oct 29.


Conjugates of adenosine mimics and d-arginine-rich peptides (ARCs) are potent inhibitors of protein kinases (PKs) from the AGC group. Labeling ARCs with fluorescent dyes or immobilizing on chip surfaces gives fluorescent probes (ARC-Photo) and biosensors that can be used for high-throughput screening (HTS) of inhibitors of protein kinases. The bisubstrate character (simultaneous association with both binding sites of the kinase) and high affinity of ARCs allow ARC-based probes and sensors to be used for characterization of inhibitors targeted to either binding site of the kinase with affinities in whole nanomolar to micromolar range. The ability to penetrate cell plasma membrane and bind to the target kinase fused with a fluorescent protein leads to the possibility to use ARC-Photo probes for high content screening (HCS) of inhibitors in cellular milieu with detection of intensity of Förster resonance energy transfer (FRET) between two fluorophores.

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Animals
  • Biosensing Techniques / methods*
  • Drug Evaluation, Preclinical / methods
  • Fluorescence Resonance Energy Transfer / methods*
  • Fluorescent Dyes / chemistry*
  • Fluorescent Dyes / metabolism
  • Humans
  • Peptides / chemistry*
  • Peptides / metabolism
  • Protein Array Analysis / methods*
  • Protein Binding
  • Protein Kinase Inhibitors / chemistry*
  • Protein Kinase Inhibitors / metabolism
  • Protein Kinases / chemistry*


  • Fluorescent Dyes
  • Peptides
  • Protein Kinase Inhibitors
  • Protein Kinases