Abstract
We report a microarray format for the detection of kinase functionality/inhibition based on marking peptide phosphorylation/biotinylation events by the attachment of gold nanoparticles followed by silver deposition for signal enhancement. The detection principle is resonance light scattering (RLS) or surface-enhanced Raman spectroscopy (SERS). Alpha-catalytic subunit of cyclic adenosine 5'-monophosphate (cAMP) dependent protein kinase (PKA) and its well-known substrate, kemptide, are 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, inhibition assay with inhibitors demonstrate the ability to detect kinase inhibition as well as derive IC50 (half maximal inhibitory concentration) plots.
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