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Title: Molecular Basis for the Association of Human E4B U Box Ubiquitin Ligase with E2-Conjugating Enzymes UbcH5c and Ubc4

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.

Authors:
; ; ; ; ; ; ; ; ;  [1];  [2];  [2]
  1. (Hokkaido)
  2. (
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
USDOE
OSTI Identifier:
1002670
Resource Type:
Journal Article
Resource Relation:
Journal Name: Structure; Journal Volume: 18; Journal Issue: (8) ; 08, 2010
Country of Publication:
United States
Language:
ENGLISH
Subject:
08 HYDROGEN; CALORIMETRY; CHAINS; CRYSTALLOGRAPHY; ELONGATION; ENZYMES; HYDROGEN; LIGASES; MACHINERY; MONOMERS; PROTEINS; REGULATIONS; SCALARS; SPECTROSCOPY

Citation Formats

Benirschke, Robert C., Thompson, James R., Nominé, Yves, Wasielewski, Emeric, Juranić, Nenad, Macura, Slobodan, Hatakeyama, Shigetsugu, Nakayama, Keiichi I., Botuyan, Maria Victoria, Mer, Georges, Mayo), and Kyushu). Molecular Basis for the Association of Human E4B U Box Ubiquitin Ligase with E2-Conjugating Enzymes UbcH5c and Ubc4. United States: N. p., 2010. Web. doi:10.1016/j.str.2010.04.017.
Benirschke, Robert C., Thompson, James R., Nominé, Yves, Wasielewski, Emeric, Juranić, Nenad, Macura, Slobodan, Hatakeyama, Shigetsugu, Nakayama, Keiichi I., Botuyan, Maria Victoria, Mer, Georges, Mayo), & Kyushu). Molecular Basis for the Association of Human E4B U Box Ubiquitin Ligase with E2-Conjugating Enzymes UbcH5c and Ubc4. United States. doi:10.1016/j.str.2010.04.017.
Benirschke, Robert C., Thompson, James R., Nominé, Yves, Wasielewski, Emeric, Juranić, Nenad, Macura, Slobodan, Hatakeyama, Shigetsugu, Nakayama, Keiichi I., Botuyan, Maria Victoria, Mer, Georges, Mayo), and Kyushu). Tue . "Molecular Basis for the Association of Human E4B U Box Ubiquitin Ligase with E2-Conjugating Enzymes UbcH5c and Ubc4". United States. doi:10.1016/j.str.2010.04.017.
@article{osti_1002670,
title = {Molecular Basis for the Association of Human E4B U Box Ubiquitin Ligase with E2-Conjugating Enzymes UbcH5c and Ubc4},
author = {Benirschke, Robert C. and Thompson, James R. and Nominé, Yves and Wasielewski, Emeric and Juranić, Nenad and Macura, Slobodan and Hatakeyama, Shigetsugu and Nakayama, Keiichi I. and Botuyan, Maria Victoria and Mer, Georges and Mayo) and Kyushu)},
abstractNote = {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.},
doi = {10.1016/j.str.2010.04.017},
journal = {Structure},
number = (8) ; 08, 2010,
volume = 18,
place = {United States},
year = {Tue Sep 07 00:00:00 EDT 2010},
month = {Tue Sep 07 00:00:00 EDT 2010}
}