title: Sensitive detection of protein kinase A activity in cell lysates by peptide microarray-based assay.
abstract: 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.
authors.affiliation: State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, P R China.
authors.forename: Tao
authors.initials: T
authors.lastname: Li
authors.forename: Xia
authors.initials: X
authors.lastname: Liu
authors.forename: Dianjun
authors.initials: D
authors.lastname: Liu
authors.forename: Zhenxin
authors.initials: Z
authors.lastname: Wang
mesh_terms: D000595:Amino Acid Sequence; D016923:Cell Death; D045744:Cell Line, Tumor; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D057075:Enzyme Assays / Q000379:methods*; D006801:Humans; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D010766:Phosphorylation; D040081:Protein Array Analysis / Q000379:methods*
chemical_list: D010455:Peptides; D017868:Cyclic AMP-Dependent Protein Kinases