Crystal Structure of a Ube2S-Ubiquitin Conjugate

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 “...

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Main Authors:	Lorenz, Sonja, Bhattacharyya, Moitrayee, Feiler, Christian, Rape, Michael, Kuriyan, John
Format:	Online
Language:	English
Published:	Public Library of Science 2016
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4734694/

id	pubmed-4734694
recordtype	oai_dc
spelling	pubmed-47346942016-02-04 Crystal Structure of a Ube2S-Ubiquitin Conjugate Lorenz, Sonja Bhattacharyya, Moitrayee Feiler, Christian Rape, Michael Kuriyan, John Research Article 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. Public Library of Science 2016-02-01 /pmc/articles/PMC4734694/ /pubmed/26828794 http://dx.doi.org/10.1371/journal.pone.0147550 Text en © 2016 Lorenz et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Lorenz, Sonja Bhattacharyya, Moitrayee Feiler, Christian Rape, Michael Kuriyan, John
spellingShingle	Lorenz, Sonja Bhattacharyya, Moitrayee Feiler, Christian Rape, Michael Kuriyan, John Crystal Structure of a Ube2S-Ubiquitin Conjugate
author_facet	Lorenz, Sonja Bhattacharyya, Moitrayee Feiler, Christian Rape, Michael Kuriyan, John
author_sort	Lorenz, Sonja
title	Crystal Structure of a Ube2S-Ubiquitin Conjugate
title_short	Crystal Structure of a Ube2S-Ubiquitin Conjugate
title_full	Crystal Structure of a Ube2S-Ubiquitin Conjugate
title_fullStr	Crystal Structure of a Ube2S-Ubiquitin Conjugate
title_full_unstemmed	Crystal Structure of a Ube2S-Ubiquitin Conjugate
title_sort	crystal structure of a ube2s-ubiquitin conjugate
description	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.
publisher	Public Library of Science
publishDate	2016
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4734694/
_version_	1613531106748923904

Crystal Structure of a Ube2S-Ubiquitin Conjugate

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