Sensitive Detection of Protein Kinase A Activity in Cell Lysates by Peptide Microarray-Based Assay

摘要

In the present work, the activities of protein kinase A (PKA) in cell lysates have been detected by a peptide microarray-based resonance light scattering assay with gold nanoparticle probes. Highly sensitive detection of PKA activity in 0.1 μg total cell proteins of SHG-44 cell lysate (corresponding to 200 cells) is achieved by a selected peptide substrate. The experimental results also demonstrate that the assay can be employed to evaluate expression levels of PKA activity in different cell lines and chemical (e.g., Forskolin )-mediated PKA activity fluctuation in living cells. In addition, PKA inhibition by the inhibitor (H89) is shown, suggesting the potential for screening PKA inhibitors at the living cell level.