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A disulphide bond in the E2 enzyme Pex4p modulates ubiquitin-conjugating activity

The ubiquitin-conjugating enzyme Pex4p together with its binding partner, the peroxisomal membrane protein Pex22p, co-ordinates cysteine-dependent ubiquitination of the cycling receptor protein Pex5p. Unusually for an ubiquitin-conjugating enzyme, Saccharomyces cerevisiae Pex4p can form a disulphide...

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Autores principales: Williams, Chris, van den Berg, Marlene, Stanley, Will A., Wilmanns, Matthias, Distel, Ben
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6505396/
https://www.ncbi.nlm.nih.gov/pubmed/23896733
http://dx.doi.org/10.1038/srep02212
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author Williams, Chris
van den Berg, Marlene
Stanley, Will A.
Wilmanns, Matthias
Distel, Ben
author_facet Williams, Chris
van den Berg, Marlene
Stanley, Will A.
Wilmanns, Matthias
Distel, Ben
author_sort Williams, Chris
collection PubMed
description The ubiquitin-conjugating enzyme Pex4p together with its binding partner, the peroxisomal membrane protein Pex22p, co-ordinates cysteine-dependent ubiquitination of the cycling receptor protein Pex5p. Unusually for an ubiquitin-conjugating enzyme, Saccharomyces cerevisiae Pex4p can form a disulphide bond between the cysteine residues at positions 105 and 146. We found that mutating the disulphide forming cysteine residues in Pex4p to serines does not disturb the secondary structure of the protein but does reduce the in vitro activity of Pex4p. From the crystal structure of Pex4p C105S, C146S in complex with the soluble domain of Pex22p, we observe a narrowing of the active site cleft, caused by loss of the disulphide bond. This modification of the active site microenvironment is likely to restrict access of ubiquitin to the active site cysteine, modulating Pex4p activity. Finally, based on sequence and structural alignments, we have identified other ubiquitin-conjugating enzymes that may contain disulphide bonds.
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spelling pubmed-65053962019-05-21 A disulphide bond in the E2 enzyme Pex4p modulates ubiquitin-conjugating activity Williams, Chris van den Berg, Marlene Stanley, Will A. Wilmanns, Matthias Distel, Ben Sci Rep Article The ubiquitin-conjugating enzyme Pex4p together with its binding partner, the peroxisomal membrane protein Pex22p, co-ordinates cysteine-dependent ubiquitination of the cycling receptor protein Pex5p. Unusually for an ubiquitin-conjugating enzyme, Saccharomyces cerevisiae Pex4p can form a disulphide bond between the cysteine residues at positions 105 and 146. We found that mutating the disulphide forming cysteine residues in Pex4p to serines does not disturb the secondary structure of the protein but does reduce the in vitro activity of Pex4p. From the crystal structure of Pex4p C105S, C146S in complex with the soluble domain of Pex22p, we observe a narrowing of the active site cleft, caused by loss of the disulphide bond. This modification of the active site microenvironment is likely to restrict access of ubiquitin to the active site cysteine, modulating Pex4p activity. Finally, based on sequence and structural alignments, we have identified other ubiquitin-conjugating enzymes that may contain disulphide bonds. Nature Publishing Group 2013-07-30 /pmc/articles/PMC6505396/ /pubmed/23896733 http://dx.doi.org/10.1038/srep02212 Text en Copyright © 2013, Macmillan Publishers Limited. All rights reserved http://creativecommons.org/licenses/by-nc-nd/3.0/ This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/3.0/
spellingShingle Article
Williams, Chris
van den Berg, Marlene
Stanley, Will A.
Wilmanns, Matthias
Distel, Ben
A disulphide bond in the E2 enzyme Pex4p modulates ubiquitin-conjugating activity
title A disulphide bond in the E2 enzyme Pex4p modulates ubiquitin-conjugating activity
title_full A disulphide bond in the E2 enzyme Pex4p modulates ubiquitin-conjugating activity
title_fullStr A disulphide bond in the E2 enzyme Pex4p modulates ubiquitin-conjugating activity
title_full_unstemmed A disulphide bond in the E2 enzyme Pex4p modulates ubiquitin-conjugating activity
title_short A disulphide bond in the E2 enzyme Pex4p modulates ubiquitin-conjugating activity
title_sort disulphide bond in the e2 enzyme pex4p modulates ubiquitin-conjugating activity
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6505396/
https://www.ncbi.nlm.nih.gov/pubmed/23896733
http://dx.doi.org/10.1038/srep02212
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