| An in vitro assay for Cdc20-dependent mitotic anaphase-promoting complex activity from budding yeast. | |
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MedLine Citation:
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PMID: 19475395 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Cell cycle transitions are controlled, in part, by ubiquitin-dependent proteolysis. In mitosis, the metaphase to anaphase transition is governed by an E3 ubiquitin ligase called the cyclosome or Anaphase-Promoting Complex (APC), and a WD40-repeat protein co-factor called Cdc20. In vitro Cdc20-dependent APC (APC(Cdc20)) assays have been useful in the identification and validation of target substrates, and in the study of APC enzymology and regulation. Many aspects of the regulation of cell cycle progression have been discovered in the budding yeast Saccharomyces cerevisiae, and proteins purified from this model organism have been employed in a wide variety of in vitro assays. Here we outline a quantitative in vitro mitotic APC(Cdc20) assay that makes use of a highly active form of the APC that is purified from budding yeast cells arrested in mitosis. |
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Authors:
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Scott C Schuyler; Andrew W Murray |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural |
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: 271-85 Citation Subset: IM |
Affiliation:
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Department of Molecular and Cellular Biology, Harvard University, Cambridge, MA, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Anaphase
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physiology* Cell Cycle Proteins / physiology* Mitosis / physiology* Saccharomyces cerevisiae / physiology* Saccharomyces cerevisiae Proteins / metabolism*, physiology Ubiquitin-Protein Ligase Complexes / physiology* Ubiquitin-Protein Ligases / metabolism Ubiquitination |
| Grant Support | |
ID/Acronym/Agency:
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GM 043987/GM/NIGMS NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/CDC20 protein, S cerevisiae; 0/Cell Cycle Proteins; 0/Saccharomyces cerevisiae Proteins; EC 6.3.2.19/Ubiquitin-Protein Ligase Complexes; EC 6.3.2.19/Ubiquitin-Protein Ligases; 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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