Microarray-based kinase inhibition assay by gold nanoparticle probes.
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.
Sun
Linlin
L
State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, China.
Liu
Dianjun
D
Wang
Zhenxin
Z
D000818:Animals; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000737:chemistry; D006046:Gold / Q000737:chemistry; D006801:Humans; D020128:Inhibitory Concentration 50; D015335:Molecular Probes / Q000737:chemistry; D053758:Nanoparticles / Q000737:chemistry*; D009842:Oligopeptides / Q000737:chemistry; D010766:Phosphorylation / Q000187:drug effects; D040081:Protein Array Analysis / Q000379:methods*; D047428:Protein Kinase Inhibitors / Q000302:isolation & purification* / Q000494:pharmacology; D011494:Protein Kinases / Q000187:drug effects* / Q000378:metabolism*
D015335:Molecular Probes; D009842:Oligopeptides; D047428:Protein Kinase Inhibitors; C037642:kemptide; D006046:Gold; D011494:Protein Kinases; D017868:Cyclic AMP-Dependent Protein Kinases