A peptide microarray for the detection of protein kinase activity in cell lysate.
DNA microarray enables the analysis of DNA or mRNA expression levels, but it has not been possible to completely understand life using obtained information. Consequently, protein or peptide arrays have attracted much interest. Since the development of a practical protein microarray is still far away in light of handling difficulties, the peptide microarray is a promising tool for analyzing protein functions. We have developed a peptide microarray to detect protein kinase activity in cell lysate. All substrate peptides for kinases were immobilized chemoselectively on amino-coated glass slides. After phosphorylation of the immobilized peptides, phosphorylation was detected by fluorescence imaging. We detected the protein kinase activities, including that in cell lysate, in response to drug stimulation. Therefore, this peptide microarray would be useful for a high-throughput kinase assay of intracellular signals and would be applicable to drug screening.
Shigaki
Syuhei
S
Department of Applied Chemistry, Faculty of Engineering, Kyushu University, Fukuoka, Japan.
Yamaji
Takayuki
T
Han
Xiaoming
X
Yamanouchi
Go
G
Sonoda
Tatsuhiko
T
Okitsu
Osamu
O
Mori
Takeshi
T
Niidome
Takuro
T
Katayama
Yoshiki
Y
D002457:Cell Extracts / Q000737:chemistry*; D002460:Cell Line; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D006801:Humans; D010766:Phosphorylation; D040081:Protein Array Analysis / Q000379:methods*; D011493:Protein Kinase C / Q000378:metabolism*
D002457:Cell Extracts; D017868:Cyclic AMP-Dependent Protein Kinases; D011493:Protein Kinase C