ac4010502_si_001.pdf (2.31 MB)
Sensitive Detection of Protein Kinase A Activity in Cell Lysates by Peptide Microarray-Based Assay
journal contribution
posted on 2013-08-06, 00:00 authored by Tao Li, Xia Liu, Dianjun Liu, Zhenxin WangIn 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.