Document Detail


Preparation of synchronized human cell extracts to study ubiquitination and degradation.
MedLine Citation:
PMID:  19475397     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Adam Williamson; Lingyan Jin; Michael Rape
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  545     ISSN:  1064-3745     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2009  
Date Detail:
Created Date:  2009-05-28     Completed Date:  2009-11-02     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  301-12     Citation Subset:  IM    
Affiliation:
Department of Molecular and Cell Biology, University of California at Berkeley, Berkeley, CA, USA.
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MeSH Terms
Descriptor/Qualifier:
Cell Cycle / physiology*
Cell Cycle Proteins / metabolism*
Cell Extracts*
Hela Cells
Humans
Mitosis
Ubiquitin-Protein Ligase Complexes / physiology*
Ubiquitination*
Chemical
Reg. No./Substance:
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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