title: E1-catalyzed ubiquitin C-terminal amidation for the facile synthesis of deubiquitinase substrates.
abstract: Will Ub my partner? The ubiquitin (Ub)-activating enzyme (E1) was used to catalyze an amidation reaction to functionalize the C terminus of Ub with unique functional groups, such as thiol, azide, alkyne, and alkene groups, with high efficiency and yield (>90 %). These groups were then applied for the facile synthesis of fluorophore-conjugated ubiquitin and specifically conjugated diubiquitin substrates for deubiquitinases.
authors.affiliation: Department of Chemistry, Texas A&M University, College Station, TX 77843 (USA).
authors.forename: Xiaoyan Aria
authors.initials: XA
authors.lastname: Wang
authors.forename: Yadagiri
authors.initials: Y
authors.lastname: Kurra
authors.forename: Ying
authors.initials: Y
authors.lastname: Huang
authors.forename: Yan-Jiun
authors.initials: YJ
authors.lastname: Lee
authors.forename: Wenshe R
authors.initials: WR
authors.lastname: Liu
mesh_terms: D000475:Alkenes / Q000737:chemistry / Q000378:metabolism; D000480:Alkynes / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D001386:Azides / Q000737:chemistry / Q000378:metabolism; D055162:Biocatalysis; D007700:Kinetics; D017434:Protein Structure, Tertiary; D013379:Substrate Specificity; D013438:Sulfhydryl Compounds / Q000737:chemistry / Q000378:metabolism; D025801:Ubiquitin / Q000737:chemistry / Q000378:metabolism*; D044764:Ubiquitin-Activating Enzymes / Q000737:chemistry / Q000378:metabolism*; D064570:Ubiquitin-Specific Proteases / Q000737:chemistry / Q000378:metabolism*; D054875:Ubiquitination
chemical_list: D000475:Alkenes; D000480:Alkynes; D001386:Azides; D013438:Sulfhydryl Compounds; D025801:Ubiquitin; D064570:Ubiquitin-Specific Proteases; D044764:Ubiquitin-Activating Enzymes
keywords: deubiquitinases; diubiquitin; protein engineering; synthetic biology; ubiquitin functionalization