title: Crystal Structure of a Ube2S-Ubiquitin Conjugate.
abstract: Protein ubiquitination occurs through the sequential formation and reorganization of specific protein-protein interfaces. Ubiquitin-conjugating (E2) enzymes, such as Ube2S, catalyze the formation of an isopeptide linkage between the C-terminus of a "donor" ubiquitin and a primary amino group of an "acceptor" ubiquitin molecule. This reaction involves an intermediate, in which the C-terminus of the donor ubiquitin is thioester-bound to the active site cysteine of the E2 and a functionally important interface is formed between the two proteins. A docked model of a Ube2S-donor ubiquitin complex was generated previously, based on chemical shift mapping by NMR, and predicted contacts were validated in functional studies. We now present the crystal structure of a covalent Ube2S-ubiquitin complex. The structure contains an interface between Ube2S and ubiquitin in trans that resembles the earlier model in general terms, but differs in detail. The crystallographic interface is more hydrophobic than the earlier model and is stable in molecular dynamics (MD) simulations. Remarkably, the docked Ube2S-donor complex converges readily to the configuration seen in the crystal structure in 3 out of 8 MD trajectories. Since the crystallographic interface is fully consistent with mutational effects, this indicates that the structure provides an energetically favorable representation of the functionally critical Ube2S-donor interface.
authors.affiliation: California Institute for Quantitative Biosciences, University of California, Berkeley, California, United States of America.|Department of Molecular and Cell Biology, University of California, Berkeley, California, United States of America.|Rudolf Virchow Center for Experimental Biomedicine, University of Würzburg, Würzburg, Germany.
authors.forename: Sonja
authors.initials: S
authors.lastname: Lorenz
authors.affiliation: California Institute for Quantitative Biosciences, University of California, Berkeley, California, United States of America.|Department of Molecular and Cell Biology, University of California, Berkeley, California, United States of America.
authors.forename: Moitrayee
authors.initials: M
authors.lastname: Bhattacharyya
authors.affiliation: Rudolf Virchow Center for Experimental Biomedicine, University of Würzburg, Würzburg, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Feiler
authors.affiliation: California Institute for Quantitative Biosciences, University of California, Berkeley, California, United States of America.|Department of Molecular and Cell Biology, University of California, Berkeley, California, United States of America.|Howard Hughes Medical Institute, University of California, Berkeley, California, United States of America.
authors.forename: Michael
authors.initials: M
authors.lastname: Rape
authors.affiliation: California Institute for Quantitative Biosciences, University of California, Berkeley, California, United States of America.|Department of Molecular and Cell Biology, University of California, Berkeley, California, United States of America.|Howard Hughes Medical Institute, University of California, Berkeley, California, United States of America.|Department of Chemistry, University of California, Berkeley, California, United States of America.|Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, California, United States of America.
authors.forename: John
authors.initials: J
authors.lastname: Kuriyan
mesh_terms: D018360:Crystallography, X-Ray; D008239:Lysine / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D056004:Molecular Dynamics Simulation; D040681:Structural Homology, Protein; D025801:Ubiquitin / Q000737:chemistry; D044763:Ubiquitin-Conjugating Enzymes / Q000737:chemistry* / Q000378:metabolism
chemical_list: D025801:Ubiquitin; D044763:Ubiquitin-Conjugating Enzymes; D008239:Lysine
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