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: Contribution to journalArticle査読

    15 被引用数 (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.

    本文言語英語
    ページ(範囲)256-262
    ページ数7
    ジャーナルJournal of Biomolecular Screening
    14
    3
    DOI
    出版ステータス出版済み - 3 2009

    All Science Journal Classification (ASJC) codes

    • 分析化学
    • バイオテクノロジー
    • 生化学
    • 分子医療
    • 薬理学
    • 創薬

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