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 fun...

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Bibliographic Details
Published in:Analytical biochemistry 2008, Vol.372 (1), p.106-115
Main Authors: Han, Xiaoming, Shigaki, Syuhei, Yamaji, Takayuki, Yamanouchi, Go, Mori, Takeshi, Niidome, Takuro, Katayama, Yoshiki
Format: Article
Language:English
Subjects:
Antibody Specificity
Calibration
Cell Line, Tumor
Cell lysate
High-throughput
Humans
Indicators and Reagents - chemistry
Peptide array
Peptides - chemistry
Phosphorylation
Protein kinase
Protein Kinases - immunology
Protein Kinases - metabolism
Quantitative
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Tyrosine - metabolism
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Summary: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.
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2007.09.026