| Differential proteomic screen to evidence proteins ubiquitinated upon mitotic exit in cell-free extract of Xenopus laevis embryos. | |
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MedLine Citation:
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PMID: 18823142 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Post-translational modification of proteins via ubiquitination plays a crucial role in numerous vital functions of the cell. Polyubiquitination is one of the key regulatory processes involved in regulation of mitotic progression. Here we describe a differential proteomic screen dedicated to identification of novel proteins ubiquitinated upon mitotic exit in cell-free extract of Xenopus laevis embryo. Mutated recombinant His6-tagged ubiquitin (Ubi (K48R)) was added to mitotic extract from which we purified conjugated proteins, as well as associated proteins in nondenaturing conditions by cobalt affinity chromatography. Proteins eluted from Ubi (K48R) supplemented and control extracts were compared by LC-MS/MS analysis after monodimensional SDS-PAGE. A total of 144 proteins potentially ubiquitinated or associated with them were identified. Forty-one percent of these proteins were shown to be involved in ubiquitination and/or proteasomal degradation pathway confirming the specificity of the screen. Twelve proteins, among them ubiquitin itself, were shown to carry a "GG" or "LRGG" remnant tag indicating their direct ubiquitination. Interestingly, sequence analysis of ubiquitinated substrates carrying these tags indicated that in Xenopus cell-free embryo extract supplemented with Ubi (K48R) the majority of polyubiquitination occurred through lysine-11 specific ubiquitin chain polymerization. The potential interest in this atypical form of ubiquitination as well as usefulness of our method in analyzing atypical polyubiquitin species is discussed. |
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Authors:
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Franck Bazile; Jean-Philippe Gagné; Geneviève Mercier; Ken Sin Lo; Aude Pascal; Julian Vasilescu; Daniel Figeys; Guy G Poirier; Jacek Z Kubiak; Franck Chesnel |
Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2008-09-30 |
Journal Detail:
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Title: Journal of proteome research Volume: 7 ISSN: 1535-3893 ISO Abbreviation: J. Proteome Res. Publication Date: 2008 Nov |
Date Detail:
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Created Date: 2008-11-10 Completed Date: 2009-01-07 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 101128775 Medline TA: J Proteome Res Country: United States |
Other Details:
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Languages: eng Pagination: 4701-14 Citation Subset: IM |
Affiliation:
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CNRS UMR 6061, Institute of Genetics & Development, University of Rennes 1, Mitosis & Meiosis Group, IFR 140 GFAS, 35 043 Rennes Cedex, France. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Cell-Free System Embryo, Nonmammalian / enzymology Female Histidine / metabolism Mitosis / physiology* Proteome / chemistry, metabolism* Proteomics / methods* Recombinant Proteins / metabolism Ubiquitin / genetics, metabolism Ubiquitination* Xenopus Proteins / metabolism* Xenopus laevis |
| Chemical | |
Reg. No./Substance:
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0/Proteome; 0/Recombinant Proteins; 0/Ubiquitin; 0/Xenopus Proteins; 71-00-1/Histidine |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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