title: A quantitative peptide array for evaluation of protein kinase activity.
abstract: 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.
authors.affiliation: Graduate School of System Life Sciences, Kyushu University, Nishi-ku Fukuoka 819-0395, Japan.
authors.lastname: Xiaoming Han
authors.lastname: Syuhei Shigaki
authors.lastname: Takayuki Yamaji
authors.lastname: Go Yamanouchi
authors.lastname: Takeshi Mori
authors.lastname: Takuro Niidome
authors.lastname: Yoshiki Katayama
mesh_terms: 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
chemical_list: D007202:Indicators and Reagents; D010455:Peptides; D014443:Tyrosine; D011494:Protein Kinases