title: Catalysis of lysine 48-specific ubiquitin chain assembly by residues in E2 and ubiquitin.
abstract: Protein ubiquitination is catalyzed by ubiquitin-conjugating enzymes (E2s) in collaboration with ubiquitin-protein ligases (E3s). This process depends on nucleophilic attack by a substrate lysine on a thioester bond linking the C terminus of ubiquitin to a cysteine in the E2 active site. Different E2 family members display specificity for lysines in distinct contexts. We addressed the mechanistic basis for this lysine selectivity in Ubc1, an E2 that catalyzes the ubiquitination of lysine 48 (K48) in ubiquitin, leading to the formation of K48-linked polyubiquitin chains. We identified a cluster of polar residues near the Ubc1 active site, as well as a residue in ubiquitin itself, that are required for catalysis of K48-specific ubiquitin ligation, but not for general activity toward other lysines. Our results suggest that the active site of Ubc1, as well as the surface of ubiquitin, contains specificity determinants that channel specific lysines to the central residues involved directly in catalysis.
authors.affiliation: Department of Physiology, University of California, San Francisco, San Francisco, CA 94158, USA.
authors.forename: Monica C
authors.initials: MC
authors.lastname: Rodrigo-Brenni
authors.forename: Scott A
authors.initials: SA
authors.lastname: Foster
authors.forename: David O
authors.initials: DO
authors.lastname: Morgan
mesh_terms: D000409:Alanine; D001665:Binding Sites; D002384:Catalysis; D020134:Catalytic Domain; D005973:Glutamine; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D007930:Leucine; D008239:Lysine; D008958:Models, Molecular; D025821:Polyubiquitin / Q000378:metabolism; D011487:Protein Conformation; D011499:Protein Processing, Post-Translational; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013329:Structure-Activity Relationship; D013912:Threonine; D014443:Tyrosine; D025801:Ubiquitin / Q000378:metabolism*; D044763:Ubiquitin-Conjugating Enzymes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D054875:Ubiquitination
chemical_list: D029701:Saccharomyces cerevisiae Proteins; D025801:Ubiquitin; D005973:Glutamine; D025821:Polyubiquitin; D013912:Threonine; D014443:Tyrosine; C066752:UBC1 protein, S cerevisiae; D044763:Ubiquitin-Conjugating Enzymes; D007930:Leucine; D008239:Lysine; D000409:Alanine
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