CDK-activating kinases: Detection and activity measurements Journal Article

  


Authors: Larochelle, S.; Fisher, R. P.
Article Title: CDK-activating kinases: Detection and activity measurements
Abstract: All cyclin-dependent kinases (CDKs) involved in eukaryotic cell cycle control require phosphorylation at a conserved threonine (or serine) residue within the activation- or T-loop to attain full enzymatic activity. The enzyme responsible for this activating phosphorylation, the CDK-activating kinase (CAK), is therefore essential for proliferation of all eukaryotic cells. We describe methods to assess the T-loop phosphorylation state of the major mammalian CDKs in vivo; to measure the levels of CAK activity in cell-free extracts; and to analyze the abundance, subunit composition, and phosphorylation state of CAK complexes in metazoan cells. When derangement of normal CDK regulation is suspected as a cause of disturbed cell cycle progression, the combination of these methodologies can ascertain whether a primary CAK defect is the explanation.
Keywords: mouse; animal; metabolism; animals; mice; cell cycle; in vitro study; phosphorylation; physiology; monoclonal antibody; immunology; antibodies, monoclonal; chemistry; immunoprecipitation; immunoblotting; binding site; binding sites; cyclin-dependent kinases; cyclin dependent kinase; gel mobility shift assay; electrophoretic mobility shift assay; polyacrylamide gel electrophoresis; electrophoresis, polyacrylamide gel; cyclin dependent kinase activating kinase; cyclin-dependent kinase-activating kinase
Journal Title: Methods in Molecular Biology
Volume: 296
ISSN: 1064-3745
Publisher: Humana Press Inc  
Date Published: 2005-01-01
Start Page: 279
End Page: 290
Language: English
PUBMED: 15576939
PROVIDER: scopus
DOI: 10.1385/1-59259-857-9:279
DOI/URL:

http://www.scopus.com/inward/record.url?eid=2-s2.0-16644375488&partnerID=40&md5=c4fa4548c407d4f22e9c629c824e93a1
Notes: Chapter 16 in "Cell Cycle Control: Mechanisms and Protocols" (ISBN: 978-1-58829-144-8) --- Cited By (since 1996): 1 - Export Date: 24 October 2012 - Source: Scopus
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MSK Authors
  1. Robert P Fisher
    28 Fisher
  2. Stephane Larochelle
    10 Larochelle
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