Expression and assay of HECT domain ligases.

Beaudenon, Sylvie; Dastur, Anahita; Huibregtse, Jon M

Beaudenon, Sylvie; Dastur, Anahita; Huibregtse, Jon M.

Affiliation

Beaudenon S; Institute for Cellular and Molecular Biology, University of Texas at Austin, Austin, TX 78712-1095, USA.

Methods Enzymol ; 398: 112-25, 2005.

Article in En | MEDLINE | ID: mdl-16275324

ABSTRACT

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.

Subject(s)

Ubiquitin-Protein Ligases/biosynthesis; Ubiquitin-Protein Ligases/genetics; Endosomal Sorting Complexes Required for Transport; Humans; Protein Structure, Tertiary; Saccharomyces cerevisiae/enzymology; Saccharomyces cerevisiae/genetics; Saccharomyces cerevisiae Proteins/analysis; Saccharomyces cerevisiae Proteins/biosynthesis; Saccharomyces cerevisiae Proteins/genetics; Ubiquitin-Conjugating Enzymes/metabolism; Ubiquitin-Protein Ligase Complexes/analysis; Ubiquitin-Protein Ligase Complexes/biosynthesis; Ubiquitin-Protein Ligase Complexes/genetics; Ubiquitin-Protein Ligases/analysis

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Collection: 01-internacional Database: MEDLINE Main subject: Ubiquitin-Protein Ligases Limits: Humans Language: En Journal: Methods Enzymol Year: 2005 Document type: Article Affiliation country: United States

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