Responsive microsecond-lifetime photoluminescent probes for analysis of protein kinases and their inhibitors

Biochim Biophys Acta. 2013 Jul;1834(7):1330-5. doi: 10.1016/j.bbapap.2013.02.039. Epub 2013 Mar 14.

Abstract

Responsive ARC-Lum probes were used for measurement of the concentration of active protein kinases (PKs) and determination of affinity of inhibitors of PKs. ARC-Lum probes incorporate thiophene or a selenophene heterocycle and a fluorophore conjugated to the lysine residue in the peptide fragment. In the complex with a PK, ARC-Lum probes emit long-lifetime (microsecond-scale) luminescence at the emission wavelengths of the fluorescent label if the complex is illuminated at the excitation wavelength of the thiophene- or selenophene-containing phosphorescence donors. Bisubstrate ARC-Lum probes bind with sub-nanomolar affinity with several PKs of the AGC group. This article is part of a Special Issue entitled: Inhibitors of Protein Kinases (2012).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algorithms
  • Anisotropy
  • Binding Sites
  • Binding, Competitive
  • Biocatalysis / drug effects
  • Fluorescent Dyes / chemistry
  • Fluorescent Dyes / metabolism*
  • Kinetics
  • Lysine / chemistry
  • Lysine / metabolism*
  • Models, Chemical
  • Molecular Structure
  • Protein Binding
  • Protein Kinase C / antagonists & inhibitors
  • Protein Kinase C / chemistry
  • Protein Kinase C / metabolism
  • Protein Kinase Inhibitors / metabolism*
  • Protein Kinase Inhibitors / pharmacology
  • Protein Kinases / chemistry
  • Protein Kinases / metabolism*
  • Spectrophotometry
  • Staurosporine / metabolism
  • Staurosporine / pharmacology
  • Thiophenes / chemistry
  • Thiophenes / metabolism
  • Time Factors

Substances

  • Fluorescent Dyes
  • Protein Kinase Inhibitors
  • Thiophenes
  • Protein Kinases
  • Protein Kinase C
  • Staurosporine
  • Lysine