The Peptide Microarray-Based Resonance Light Scattering Assay for Sensitively Detecting Intracellular Kinase Activity.
The peptide microarray technology is a robust, reliable, and efficient technique for large-scale determination of enzyme activities, and high-throughput profiling of substrate/inhibitor specificities of enzymes. Here, the activities of cyclic adenosine monophosphate (cAMP)-dependent protein kinase A (PKA) in different cell lysates have been detected by a peptide microarray-based resonance light scattering (RLS) assay with gold nanoparticle (GNP) probes. Highly sensitive detection of PKA activity in 0.1 μg total cell proteins of SHG-44 (human glioma cell) cell lysate (corresponding to 200 cells) is achieved by a selected peptide substrate. The experimental results also demonstrate that the RLS assay can be employed to evaluate the chemical regulation of intracellular kinase activity.
State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, 130022, China.
Tao
T
Li
State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, 130022, China.
Xia
X
Liu
State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, 130022, China.
Dianjun
D
Liu
State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, 130022, China. wangzx@ciac.ac.cn.
Zhenxin
Z
Wang
D000818:Animals; D045744:Cell Line, Tumor; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D006046:Gold / Q000737:chemistry; D006801:Humans; D042541:Intracellular Space / Q000201:enzymology*; D008027:Light; D057230:Limit of Detection; D053768:Metal Nanoparticles / Q000737:chemistry; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D040081:Protein Array Analysis / Q000379:methods*; D051381:Rats; D012542:Scattering, Radiation
D010455:Peptides; D006046:Gold; D017868:Cyclic AMP-Dependent Protein Kinases
Chemical regulation; Gold nanoparticle; Intracellular kinase activity; Peptide microarray; Resonance light scattering