2P-15 PDF
Identification of putative PKA substrates by quantitative phosphoproteomics with a PKA inhibitor
Protein kinase A (PKA or cAMP-dependent protein kinase) is a serine/threonine kinase that plays essential roles in the regulation of proliferation, differentiation and apoptosis. To identify kinase target substrates in a high-throughput manner, we quantified the change of phosphoproteome in the cells of which PKA activity was suppressed by H89, a PKA inhibitor. Then, we developed a computational approach to rank the potential substrates based on their primary sequences. Finally, the primary-sequence-based scoring was combined with the quantitative changes in phosphorylation to identify putative PKA substrates in living cells.