Monitoring protein kinase activity in cell lysates using a high-density peptide microarray

Xiaoming Han, Go Yamanouchi, Takeshi Mori, Jeong Hun Kang, Takuro Niidome, Yoshiki Katayama

研究成果: ジャーナルへの寄稿記事

15 引用 (Scopus)

抄録

Monitoring and targeting protein kinases is widely accepted as a promising approach for disease diagnosis and drug discovery. For this purpose, the authors have developed an original type of peptide array as a high-throughput screening assay for quantitatively evaluating kinase activity. A volume of 2 nL of peptide solution was spotted onto a formyl group-modified glass slide by using an arrayer, which was designed for use with protein chip technology. The phosphorylation was recognized by fluorescence-label antibody and detected with an automatic microarray scanner widely used in DNA chip technology. The system needs low sample volume, provides a high-density peptide array, and supplies high reproducibility. It provided enough sensitivity for inhibitor screening, even though a relatively low concentration of purified kinase was employed. The assay also proved useful for the detection of intracellular kinase activity as well as for the measurement of the fluctuations of intracellular protein kinase activity with drug stimulation. Thus, this peptide array would be applicable for kinase-targeted diagnosis, cell-based drug screening, and signal pathway investigation.

元の言語英語
ページ(範囲)256-262
ページ数7
ジャーナルJournal of Biomolecular Screening
14
発行部数3
DOI
出版物ステータス出版済み - 3 1 2009

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Microarrays
Protein Kinases
Phosphotransferases
Screening
Peptides
Monitoring
Assays
High-Throughput Screening Assays
Technology
Protein Array Analysis
Preclinical Drug Evaluations
Phosphorylation
Drug Discovery
Oligonucleotide Array Sequence Analysis
Pharmaceutical Preparations
Glass
Labels
Signal Transduction
Fluorescence
Throughput

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biotechnology
  • Biochemistry
  • Molecular Medicine
  • Pharmacology
  • Drug Discovery

これを引用

Monitoring protein kinase activity in cell lysates using a high-density peptide microarray. / Han, Xiaoming; Yamanouchi, Go; Mori, Takeshi; Kang, Jeong Hun; Niidome, Takuro; Katayama, Yoshiki.

:: Journal of Biomolecular Screening, 巻 14, 番号 3, 01.03.2009, p. 256-262.

研究成果: ジャーナルへの寄稿記事

Han, Xiaoming ; Yamanouchi, Go ; Mori, Takeshi ; Kang, Jeong Hun ; Niidome, Takuro ; Katayama, Yoshiki. / Monitoring protein kinase activity in cell lysates using a high-density peptide microarray. :: Journal of Biomolecular Screening. 2009 ; 巻 14, 番号 3. pp. 256-262.
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