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Active site alanine mutations convert deubiquitinases into high-affinity ubiquitin-binding proteins
ME. Morrow, MT. Morgan, M. Clerici, K. Growkova, M. Yan, D. Komander, TK. Sixma, M. Simicek, C. Wolberger,
Jazyk angličtina Země Anglie, Velká Británie
Typ dokumentu časopisecké články, Research Support, N.I.H., Extramural, práce podpořená grantem, Research Support, U.S. Gov't, Non-P.H.S.
Grantová podpora
309756
European Research Council - International
R01 GM095822
NIGMS NIH HHS - United States
R01 GM109102
NIGMS NIH HHS - United States
MC_U105192732
Medical Research Council - United Kingdom
U105192732
Medical Research Council - United Kingdom
P41 GM103393
NIGMS NIH HHS - United States
NLK
Free Medical Journals
od 2000 do Před 1 rokem
PubMed Central
od 2000
Europe PubMed Central
od 2000 do Před 1 rokem
Open Access Digital Library
od 2000-07-01
Medline Complete (EBSCOhost)
od 2000-07-01 do Před 1 rokem
Wiley Free Content
od 2000 do Před 1 rokem
PubMed
30150323
DOI
10.15252/embr.201745680
Knihovny.cz E-zdroje
- MeSH
- alanin genetika MeSH
- cystein genetika MeSH
- deubikvitinasy chemie genetika MeSH
- endopeptidasy chemie genetika MeSH
- katalýza MeSH
- konformace proteinů MeSH
- lidé MeSH
- mutace genetika MeSH
- Saccharomyces cerevisiae - proteiny chemie genetika MeSH
- Saccharomyces cerevisiae genetika MeSH
- specifické proteázy ubikvitinu chemie genetika MeSH
- substituce aminokyselin genetika MeSH
- trans-aktivátory chemie genetika MeSH
- transportní proteiny chemie genetika MeSH
- ubikvitin chemie genetika MeSH
- ubikvitinace genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
A common strategy for exploring the biological roles of deubiquitinating enzymes (DUBs) in different pathways is to study the effects of replacing the wild-type DUB with a catalytically inactive mutant in cells. We report here that a commonly studied DUB mutation, in which the catalytic cysteine is replaced with alanine, can dramatically increase the affinity of some DUBs for ubiquitin. Overexpression of these tight-binding mutants thus has the potential to sequester cellular pools of monoubiquitin and ubiquitin chains. As a result, cells expressing these mutants may display unpredictable dominant negative physiological effects that are not related to loss of DUB activity. The structure of the SAGA DUB module bound to free ubiquitin reveals the structural basis for the 30-fold higher affinity of Ubp8C146A for ubiquitin. We show that an alternative option, substituting the active site cysteine with arginine, can inactivate DUBs while also decreasing the affinity for ubiquitin.
Division of Biochemistry and Oncode Institute Netherlands Cancer Institute Amsterdam The Netherlands
Faculty of Medicine University of Ostrava Ostrava Czech Republic
Medical Research Council Laboratory of Molecular Biology Cambridge UK
Citace poskytuje Crossref.org
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