Document Detail


Oligomeric structure and catalytic activity of G type casein kinase. Isolation of the two subunits and renaturation experiments.
MedLine Citation:
PMID:  6571836     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Casein kinase G purified from bovine tissue is an oligomeric cyclic nucleotide-independent protein kinase made of two different monomers, namely an alpha (Mr = 38 kilodaltons) and a self-phosphorylatable beta (Mr = 27 kilodaltons) subunit. Treatment of the native enzyme under denaturing conditions (0.5 M NaCl, 4 M LiCl, and 20 to 35% formamide) resulted in a progressive selective removal of the beta subunit following gel filtration and a correlated loss of activity of the corresponding renatured enzyme. Mild digestion with papain resulted in a proteolytic alteration limited to the beta monomer with a concomitant partial loss of the enzyme activity. Isolation of the alpha and beta casein kinase G subunits was achieved by preparative reversed polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Renaturation of the proteins following sodium dodecyl sulfate removal by acetone and/or Triton X-100 treatment allowed reconstitution of a functional casein kinase G. Whereas the isolated alpha subunit was found to exhibit a weak catalytic activity, addition of the beta subunit was required for recovery of a maximal casein kinase activity. The process was dose-dependent and reached a plateau for an alpha:beta subunit molar ratio of approximately 1 to 1. These data suggest that while the casein kinase G alpha subunit bears the catalytic site, stoichiometric combination with the beta subunit is required for optimal enzymatic activity. A possible role of the beta subunit as a regulatory component of casein kinase G activity in the intact cell remains to be examined.
Authors:
C Cochet; E M Chambaz
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  258     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  1983 Feb 
Date Detail:
Created Date:  1983-03-17     Completed Date:  1983-03-17     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1403-6     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Animals
Casein Kinases
Cattle
Kinetics
Lung / enzymology*
Macromolecular Substances
Molecular Weight
Papain
Protein Kinases / isolation & purification,  metabolism*
Chemical
Reg. No./Substance:
0/Macromolecular Substances; EC 2.7.-/Protein Kinases; EC 2.7.11.1/Casein Kinases; EC 3.4.22.2/Papain

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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