title: Intrinsic disorder drives N-terminal ubiquitination by Ube2w.
abstract: Ubiquitination of the αN-terminus of protein substrates has been reported sporadically since the early 1980s. However, the identity of an enzyme responsible for this unique ubiquitin (Ub) modification has only recently been elucidated. We show the Ub-conjugating enzyme (E2) Ube2w uses a unique mechanism to facilitate the specific ubiquitination of the α-amino group of its substrates that involves recognition of backbone atoms of intrinsically disordered N termini. We present the NMR-based solution ensemble of full-length Ube2w that reveals a structural architecture unlike that of any other E2 in which its C terminus is partly disordered and flexible to accommodate variable substrate N termini. Flexibility of the substrate is critical for recognition by Ube2w, and either point mutations in or the removal of the flexible C terminus of Ube2w inhibits substrate binding and modification. Mechanistic insights reported here provide guiding principles for future efforts to define the N-terminal ubiquitome in cells.
authors.affiliation: Department of Biochemistry, University of Washington, Seattle, Washington, USA.
authors.forename: Vinayak
authors.initials: V
authors.lastname: Vittal
authors.affiliation: 1] Department of Biochemistry, University of Washington, Seattle, Washington, USA. [2] Howard Hughes Medical Institute, University of Washington, Seattle, Washington, USA.
authors.forename: Lei
authors.initials: L
authors.lastname: Shi
authors.affiliation: Department of Biochemistry, University of Washington, Seattle, Washington, USA.
authors.forename: Dawn M
authors.initials: DM
authors.lastname: Wenzel
authors.affiliation: 1] Department of Neurology, University of Michigan Medical School, Ann Arbor, Michigan, USA. [2] Department of Biochemistry, Medical College of Wisconsin, Milwaukee, Wisconsin, USA. [3] Neuroscience Research Center, Medical College of Wisconsin, Milwaukee, Wisconsin, USA.
authors.forename: K Matthew
authors.initials: KM
authors.lastname: Scaglione
authors.affiliation: Department of Biochemistry, University of Washington, Seattle, Washington, USA.
authors.forename: Emily D
authors.initials: ED
authors.lastname: Duncan
authors.affiliation: Department of Pathology, University of Michigan, Ann Arbor, Michigan, USA.
authors.forename: Venkatesha
authors.initials: V
authors.lastname: Basrur
authors.affiliation: Department of Pathology, University of Michigan, Ann Arbor, Michigan, USA.
authors.forename: Kojo S J
authors.initials: KS
authors.lastname: Elenitoba-Johnson
authors.affiliation: 1] Department of Biochemistry, University of Washington, Seattle, Washington, USA. [2] Howard Hughes Medical Institute, University of Washington, Seattle, Washington, USA.
authors.forename: David
authors.initials: D
authors.lastname: Baker
authors.affiliation: Department of Neurology, University of Michigan Medical School, Ann Arbor, Michigan, USA.
authors.forename: Henry L
authors.initials: HL
authors.lastname: Paulson
authors.affiliation: Department of Biochemistry, University of Washington, Seattle, Washington, USA.
authors.forename: Peter S
authors.initials: PS
authors.lastname: Brzovic
authors.affiliation: Department of Biochemistry, University of Washington, Seattle, Washington, USA.
authors.forename: Rachel E
authors.initials: RE
authors.lastname: Klevit
mesh_terms: D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D017354:Point Mutation / Q000235:genetics; D011485:Protein Binding; D011487:Protein Conformation; D013379:Substrate Specificity; D014451:Ubiquinone / Q000378:metabolism; D044763:Ubiquitin-Conjugating Enzymes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D054875:Ubiquitination
chemical_list: D014451:Ubiquinone; C510626:UBE2W protein, human; D044763:Ubiquitin-Conjugating Enzymes