title: Expression and assay of HECT domain ligases.
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.
authors.affiliation: Institute for Cellular and Molecular Biology, University of Texas at Austin, Austin, TX 78712-1095, USA.
authors.forename: Sylvie
authors.initials: S
authors.lastname: Beaudenon
authors.forename: Anahita
authors.initials: A
authors.lastname: Dastur
authors.forename: Jon M
authors.initials: JM
authors.lastname: Huibregtse
mesh_terms: D056827:Endosomal Sorting Complexes Required for Transport; D006801:Humans; D017434:Protein Structure, Tertiary; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D044763:Ubiquitin-Conjugating Enzymes / Q000378:metabolism; D043743:Ubiquitin-Protein Ligase Complexes / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D044767:Ubiquitin-Protein Ligases / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics*
chemical_list: D056827:Endosomal Sorting Complexes Required for Transport; D029701:Saccharomyces cerevisiae Proteins; D044763:Ubiquitin-Conjugating Enzymes; D043743:Ubiquitin-Protein Ligase Complexes; C471639:UBE3A protein, human; D044767:Ubiquitin-Protein Ligases; C469287:RSP5 protein, S cerevisiae