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PMID: 20696396 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.

Molecular basis for the association of human E4B U box ubiquitin ligase with E2-conjugating enzymes UbcH5c and Ubc4.

Structure (London, England : 1993) ·Vol. 18 ·No. 8 ·2010-08-11 ·Pages 955-65

Benirschke RC, Thompson JR, Nominé Y, Wasielewski E, Juranić N, Macura S, Hatakeyama S, Nakayama KI, Botuyan MV, Mer G

Abstract

Human E4B, also called UFD2a, is a U box-containing protein that functions as an E3 ubiquitin ligase and an E4 polyubiquitin chain elongation factor. E4B is thought to participate in the proteasomal degradation of misfolded or damaged proteins through association with chaperones. The U box domain is an anchor site for E2 ubiquitin-conjugating enzymes, but little is known of the binding mechanism. Using X-ray crystallography and NMR spectroscopy, we determined the structures of E4B U box free and bound to UbcH5c and Ubc4 E2s. Whereas previously characterized U box domains are homodimeric, we show that E4B U box is a monomer stabilized by a network of hydrogen bonds identified from scalar coupling measurements. These structural studies, complemented by calorimetry- and NMR-based binding assays, suggest an allosteric regulation of UbcH5c and Ubc4 by E4B U box and provide a molecular basis to understand how the ubiquitylation machinery involving E4B assembles.

MeSH Terms
Allosteric Regulation/genetics Calorimetry Crystallography, X-Ray Humans Hydrogen Bonding Models, Molecular Nuclear Magnetic Resonance, Biomolecular Protein Conformation Tumor Suppressor Proteins/chemistry,metabolism Ubiquitin-Conjugating Enzymes/metabolism Ubiquitin-Protein Ligase Complexes/chemistry,metabolism Ubiquitin-Protein Ligases
Chemicals
Tumor Suppressor Proteins UBE2D3 protein, human Ubiquitin-Conjugating Enzymes Ubiquitin-Protein Ligase Complexes UBE4B protein, human Ubiquitin-Protein Ligases
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Benirschke Robert C
Department of Biochemistry and Molecular Biology, Mayo Clinic College of Medicine, Rochester, MN 55905, USA.
Thompson James R
Nominé Yves
Wasielewski Emeric
Juranić Nenad
Macura Slobodan
Hatakeyama Shigetsugu
Nakayama Keiichi I
Botuyan Maria Victoria
Mer Georges
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Article Info
Journal
Structure (London, England : 1993)
Abbr.
Structure
ISSN
1878-4186
DOI
10.1016/j.str.2010.04.017
Published
2010-08-11
Pages
955-65
Language
English
Region
United States
NLM ID
101087697
PMCID
PMC3005147
Subset
IM
Grants
NCI NIH HHS · CA109449 · United States
NCI NIH HHS · R01 CA132878 · United States
NCI NIH HHS · R01 CA132878-03 · United States
NCI NIH HHS · R01 CA109449 · United States
NCI NIH HHS · R01 CA109449-05 · United States
Databases
PDB
1L1X, 2KRE, 3L1Y, 3L1Z
Corrections
CommentIn
PMID: 20696387
External Links
PubMed source DOI: 10.1016/j.str.2010.04.017 PMC Full Text View journal details
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