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タイトル
  • ja Versatile assays for high throughput screening for activators or inhibitors of intracellular proteases and their cellular regulators.
作成者
    • Hayashi, Hideki
    • Cuddy, Michael
    • Shu, Vincent Chih-Wen
    • Yip, Kenneth W
    • Madiraju, Charitha
    • Diaz, Paul
    • Matsuyama, Toshifumi
    • Kaibara, Muneshige
    • Taniyama, Kohtaro
    • Vasile, Stefan
    • Sergienko, Eduard
    • Reed, John C
権利情報
  • c 2009 Hayashi et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
内容注記
  • Other BACKGROUND: Intracellular proteases constitute a class of promising drug discovery targets. Methods for high throughput screening against these targets are generally limited to in vitro biochemical assays that can suffer many technical limitations, as well as failing to capture the biological context of proteases within the cellular pathways that lead to their activation. METHODS #ENTITYSTARTX00026; FINDINGS: We describe here a versatile system for reconstituting protease activation networks in yeast and assaying the activity of these pathways using a cleavable transcription factor substrate in conjunction with reporter gene read-outs. The utility of these versatile assay components and their application for screening strategies was validated for all ten human Caspases, a family of intracellular proteases involved in cell death and inflammation, including implementation of assays for high throughput screening (HTS) of chemical libraries and functional screening of cDNA libraries. The versatility of the technology was also demonstrated for human autophagins, cysteine proteases involved in autophagy. CONCLUSIONS: Altogether, the yeast-based systems described here for monitoring activity of ectopically expressed mammalian proteases provide a fascile platform for functional genomics and chemical library screening.
  • Other identifier:PloS one, 4(10), e7655; 2009
日付
    Created2020-12-23 , Issued2009-10-30
言語
  • eng
資源識別のタイプ journal article
出版タイプ VoR
資源識別子 URI http://hdl.handle.net/10069/22646
関連
  • isIdenticalTo PMID 19876397
  • isIdenticalTo DOI https://doi.org/10.1371/journal.pone.0007655
収録誌情報
    • ISSN 1932-6203
      • PloS one 4(10), e7655
ファイル
コンテンツ更新日時 2021-02-17