Beaudenon Sylvie, Dastur Anahita, Huibregtse Jon M
Institute for Cellular and Molecular Biology, University of Texas at Austin, Austin, TX 78712-1095, USA.
Methods Enzymol. 2005;398:112-25. doi: 10.1016/S0076-6879(05)98011-7.
HECT domain ubiquitin ligases (HECT E3s), typified by human E6AP and yeast Rsp5p, are unique among the several classes of known ubiquitin ligases in that they participate directly in the chemistry of substrate ubiquitination reactions. This chapter discusses strategies for the expression of active HECT E3s and the assays that are available for analyzing E2 interaction, ubiquitin-thioester formation, and substrate ubiquitination.
HECT结构域泛素连接酶（HECT E3s），以人类E6AP和酵母Rsp5p为代表，在几类已知的泛素连接酶中独具特色，因为它们直接参与底物泛素化反应的化学过程。本章讨论了活性HECT E3s的表达策略以及可用于分析E2相互作用、泛素硫酯形成和底物泛素化的检测方法。
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