期刊
BIOCHEMICAL JOURNAL
卷 450, 期 -, 页码 55-62出版社
PORTLAND PRESS LTD
DOI: 10.1042/BJ20120572
关键词
AlphaScreen (R); GTPase; GTP competitive inhibitor; mitoxantrone; Racl
资金
- 'Programme d'Actions Integrees de Recherches sur le Carcinome Hepatocellulaire' (PAIR-CHC)
- L'Institut National du Cancer (INCa)
- ANRS (Agence Nationale de Recherche sur le Sida et les Hepatites Virales)
- Association pour la Recherche sur le Cancer (ARC)
- INCa
- Association pour la Recherche contre le Cancer (ARC)
- Conseil Regional d'Aquitaine
- Fondation Recherche Medicale
- ARC
- French Ministry of Education and Research
RhoGTPases are GDP/GTP molecular switches that control a wide variety of cellular processes, thereby contributing to many diseases, including cancer. As a consequence, there is great interest in the identification of small-molecule inhibitors of RhoGTPases. In the present paper, using the property of GTP-loaded RhoGTPases to bind to their effectors, we describe a miniaturized and robust assay to monitor Racl GTPase activation that is suitable for large-scale high-throughput screening. A pilot compound library screen revealed that the topoisomerase II poison MTX (mitoxantrone) is an inhibitor of Racl, and also inhibits RhoA and Cdc42 in vitro. We show that MTX prevents GTP binding to RhoA/Racl/Cdc42 in vitro. Furthermore, MTX strongly inhibits RhoGTPase-mediated F-actin (filamentous actin) reorganization and cell migration. Hence, we report a novel biochemical assay yielding the identification of RhoGTPase inhibitors and we present a proof-of-concept validation with the identification of MTX as a novel pan-RhoGTPase inhibitor.
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