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PMID: 20164921 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

Active site remodelling accompanies thioester bond formation in the SUMO E1.

Nature ·Vol. 463 ·No. 7283 ·2010-02-18 ·Pages 906-12

Olsen SK, Capili AD, Lu X, Tan DS, Lima CD

Abstract

E1 enzymes activate ubiquitin (Ub) and ubiquitin-like (Ubl) proteins in two steps by carboxy-terminal adenylation and thioester bond formation to a conserved catalytic cysteine in the E1 Cys domain. The structural basis for these intermediates remains unknown. Here we report crystal structures for human SUMO E1 in complex with SUMO adenylate and tetrahedral intermediate analogues at 2.45 and 2.6 A, respectively. These structures show that side chain contacts to ATP.Mg are released after adenylation to facilitate a 130 degree rotation of the Cys domain during thioester bond formation that is accompanied by remodelling of key structural elements including the helix that contains the E1 catalytic cysteine, the crossover and re-entry loops, and refolding of two helices that are required for adenylation. These changes displace side chains required for adenylation with side chains required for thioester bond formation. Mutational and biochemical analyses indicate these mechanisms are conserved in other E1s.

MeSH Terms
Adenosine Triphosphate/metabolism Amino Acid Sequence Biocatalysis Catalytic Domain/physiology Conserved Sequence Crystallography, X-Ray Cysteine/chemistry,metabolism Humans Magnesium/metabolism Models, Molecular Molecular Sequence Data Protein Conformation SUMO-1 Protein/chemistry,metabolism Saccharomyces cerevisiae Saccharomyces cerevisiae Proteins/metabolism Small Ubiquitin-Related Modifier Proteins/metabolism Sulfides/metabolism Ubiquitin/metabolism Ubiquitin-Activating Enzymes/chemistry,metabolism Ubiquitins/metabolism
Chemicals
SMT3 protein, S cerevisiae SUMO-1 Protein Saccharomyces cerevisiae Proteins Small Ubiquitin-Related Modifier Proteins Sulfides Ubiquitin Ubiquitins Adenosine Triphosphate Ubiquitin-Activating Enzymes Magnesium Cysteine
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Olsen Shaun K
Structural Biology, Sloan-Kettering Institute, New York, New York 10065, USA.
Capili Allan D
Lu Xuequan
Tan Derek S
Lima Christopher D
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Article Info
Journal
Nature
Abbr.
Nature
ISSN
1476-4687
DOI
10.1038/nature08765
Published
2010-02-18
Pages
906-12
Language
English
Region
England
NLM ID
0410462
PMCID
PMC2866016
Subset
IM
Grants
NCRR NIH HHS · P41 RR015301 · United States
NIAID NIH HHS · R01 AI068038-03 · United States
NIGMS NIH HHS · F32 GM075695-03 · United States
NIAID NIH HHS · R01 AI068038 · United States
NIGMS NIH HHS · F32 GM075695 · United States
NIGMS NIH HHS · R01 GM065872-09 · United States
NIGMS NIH HHS · R01 GM065872 · United States
NCRR NIH HHS · RR-15301 · United States
NIAID NIH HHS · R01 AI068038-02 · United States
Corrections
CommentIn
PMID: 20164915
CommentIn
PMID: 20399179
External Links
PubMed source DOI: 10.1038/nature08765 PMC Full Text View journal details
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