| Preparation of synchronized human cell extracts to study ubiquitination and degradation. | |
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MedLine Citation:
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PMID: 19475397 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Ubiquitination and protein degradation regulate cell cycle progression in all eukaryotes. During mitosis, ubiquitination by the Anaphase-Promoting Complex/Cyclosome (APC/C) triggers sister chromatid separation and mitotic exit. The APC/C is tightly regulated by phosphorylation, ubiquitination, association of activators or inhibitors, and competitive binding of substrates. Much of our understanding of the mechanism of APC/C-dependent ubiquitination has been obtained from studies using extracts of Xenopus laevis eggs or synchronized human tissue culture cells. Here, we describe protocols to prepare extracts of synchronized human cells, and discuss experiments to use extracts for the biochemical analysis of APC/C-dependent ubiquitination. |
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Authors:
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Adam Williamson; Lingyan Jin; Michael Rape |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Methods in molecular biology (Clifton, N.J.) Volume: 545 ISSN: 1064-3745 ISO Abbreviation: Methods Mol. Biol. Publication Date: 2009 |
Date Detail:
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Created Date: 2009-05-28 Completed Date: 2009-11-02 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9214969 Medline TA: Methods Mol Biol Country: United States |
Other Details:
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Languages: eng Pagination: 301-12 Citation Subset: IM |
Affiliation:
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Department of Molecular and Cell Biology, University of California at Berkeley, Berkeley, CA, USA. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Cell Cycle
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physiology* Cell Cycle Proteins / metabolism* Cell Extracts* Hela Cells Humans Mitosis Ubiquitin-Protein Ligase Complexes / physiology* Ubiquitination* |
| Chemical | |
Reg. No./Substance:
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0/Cell Cycle Proteins; 0/Cell Extracts; EC 6.3.2.19/Ubiquitin-Protein Ligase Complexes; EC 6.3.2.19/anaphase-promoting complex |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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