A quantitative peptide array for evaluation of protein kinase activity.
Peptide array, which is known as an emerging technology, has been developed for identification of protein kinase activity. For this purpose, the ability of quantitative analysis is very important because the absolute change in protein kinase activity is critical for the determination of cellular function. Here we report an original type of peptide array for quantitative evaluation of protein kinase activity by fluorescence imaging. We used the peptide array for the quantitative evaluation of the nonreceptor tyrosine kinase c-Src activity as a model for detecting protein kinase activities. By using positive and negative control peptides, we obtained the actual ratio of tyrosine phosphorylation of substrate peptide not only by purified c-Src but also by c-Src in cell lysate. In addition, the experimental approach provided simple immobilization of peptide. Our sensitive, specific, and high-throughput peptide array can be used for quantitative evaluation of kinase activity and potentially can be applied to drug discovery and screening.
Xiaoming Han
Graduate School of System Life Sciences, Kyushu University, Nishi-ku Fukuoka 819-0395, Japan.
Syuhei Shigaki
Takayuki Yamaji
Go Yamanouchi
Takeshi Mori
Takuro Niidome
Yoshiki Katayama
D000918:Antibody Specificity; D002138:Calibration; D045744:Cell Line, Tumor; D006801:Humans; D007202:Indicators and Reagents / Q000737:chemistry; D010455:Peptides / Q000737:chemistry*; D010766:Phosphorylation; D011494:Protein Kinases / Q000276:immunology / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014443:Tyrosine / Q000378:metabolism
D007202:Indicators and Reagents; D010455:Peptides; D014443:Tyrosine; D011494:Protein Kinases