A quantitative peptide array for evaluation of protein kinase activity

Xiaoming Han, Syuhei Shigaki, Takayuki Yamaji, Go Yamanouchi, Takeshi Mori, Takuro Niidome, Yoshiki Katayama

Research output: Contribution to journalArticle

25 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)106-115
Number of pages10
JournalAnalytical Biochemistry
Volume372
Issue number1
DOIs
Publication statusPublished - Jan 1 2008

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Protein Kinases
Peptides
Preclinical Drug Evaluations
Phosphorylation
Optical Imaging
Drug Discovery
Immobilization
Tyrosine
Screening
Phosphotransferases
Fluorescence
Throughput
Technology
Imaging techniques
Substrates
Chemical analysis

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

A quantitative peptide array for evaluation of protein kinase activity. / Han, Xiaoming; Shigaki, Syuhei; Yamaji, Takayuki; Yamanouchi, Go; Mori, Takeshi; Niidome, Takuro; Katayama, Yoshiki.

In: Analytical Biochemistry, Vol. 372, No. 1, 01.01.2008, p. 106-115.

Research output: Contribution to journalArticle

Han, Xiaoming ; Shigaki, Syuhei ; Yamaji, Takayuki ; Yamanouchi, Go ; Mori, Takeshi ; Niidome, Takuro ; Katayama, Yoshiki. / A quantitative peptide array for evaluation of protein kinase activity. In: Analytical Biochemistry. 2008 ; Vol. 372, No. 1. pp. 106-115.
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