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Expression and assay of HECT domain ligases
Sylvie Beaudenon1, Anahita Dastur, Jon M Huibregtse
1Institute for Cellular and Molecular Biology, University of Texas at Austin, Austin, TX 78712-1095, USA.
Methods in Enzymology
|November 9, 2005
Abstract:
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.
