Microarray-based kinase inhibition assay by gold nanoparticle probes

Anal Chem. 2007 Jan 15;79(2):773-7. doi: 10.1021/ac061687u.

Abstract

We report on the development of a new class of kinase microarray for the detection of kinase inhibition based on marking peptide phosphorylation/biotinylation events by attachment of gold nanoparticles followed by silver deposition for signal enhancement. The alpha-catalytic subunit of cyclic adenosine 5'-monophosphate-dependent protein kinase (PKA), and its well-known substrate, kemptide, were used for the purpose of monitoring phosphorylation and inhibition. As expected, highly selective inhibition of PKA is demonstrated with the four inhibitors: H89, HA1077, mallotoxin, and KN62. Furthermore, an inhibition assay demonstrates the ability to detect kinase inhibition as well as derive IC50 (half-maximal inhibitory concentration) plots.

Publication types

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

MeSH terms

  • Animals
  • Cyclic AMP-Dependent Protein Kinases / antagonists & inhibitors
  • Cyclic AMP-Dependent Protein Kinases / chemistry
  • Gold / chemistry
  • Humans
  • Inhibitory Concentration 50
  • Molecular Probes / chemistry
  • Nanoparticles / chemistry*
  • Oligopeptides / chemistry
  • Phosphorylation / drug effects
  • Protein Array Analysis / methods*
  • Protein Kinase Inhibitors / isolation & purification*
  • Protein Kinase Inhibitors / pharmacology
  • Protein Kinases / drug effects*
  • Protein Kinases / metabolism*

Substances

  • Molecular Probes
  • Oligopeptides
  • Protein Kinase Inhibitors
  • kemptide
  • Gold
  • Protein Kinases
  • Cyclic AMP-Dependent Protein Kinases