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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
27
|
| pubmed:dateCreated |
1988-10-19
|
| pubmed:abstractText |
Ca2+/calmodulin-dependent protein kinase II (CaM-kinase II) autophosphorylated under limiting conditions (7 microM [gamma-32P]ATP, 500 microM magnesium acetate, 4 degrees C) was analyzed by CNBr cleavage and peptide mapping to determine the site of autophosphorylation that brings about transition of the kinase to the Ca2+-independent form. Reverse phase high performance liquid chromatography (HPLC) (C3) revealed one major CN-Br 32P-peptide (CB1) that eluted at about 6% propanol. This peptide contained [32P]threonine, but almost no [32P]serine, and migrated as a single band (Mr = 3000-3500) in polyacrylamide gels run in the presence of urea and sodium dodecyl sulfate. The properties of CB1 were compared to the properties of a 26-residue synthetic peptide containing the CaM-binding and inhibitory domains as well as a consensus phosphorylation sequence (-Arg-Gln-Glu-Thr-) of rat brain CaM-kinase II (residues 282-307 and 283-308 of the alpha and beta subunits, respectively). CB1 and the synthetic peptide comigrated in urea/sodium dodecyl sulfate gels, co-eluted from reverse phase HPLC (C3 and C18) and from Sephadex G-50, and exhibited Ca2+-dependent calmodulin-binding properties. When the two peptides were subjected to automated Edman sequence analysis, both exhibited a burst of 32P release at cycle 5, which is consistent with the expected amino-terminal sequence of the two peptides, i.e. His-Arg-Gln-Glu-Thr(PO4)-. These findings indicate that autophosphorylation of Thr286 (alpha subunit) and Thr287 (beta subunit) is responsible for transition of CaM-kinase II to the Ca2+-independent form.
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| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Adenosine Triphosphate,
http://linkedlifedata.com/resource/pubmed/chemical/Calcium,
http://linkedlifedata.com/resource/pubmed/chemical/Calmodulin,
http://linkedlifedata.com/resource/pubmed/chemical/Cyanogen Bromide,
http://linkedlifedata.com/resource/pubmed/chemical/Peptide Fragments,
http://linkedlifedata.com/resource/pubmed/chemical/Phosphoserine,
http://linkedlifedata.com/resource/pubmed/chemical/Phosphothreonine,
http://linkedlifedata.com/resource/pubmed/chemical/Protein Kinases
|
| pubmed:status |
MEDLINE
|
| pubmed:month |
Sep
|
| pubmed:issn |
0021-9258
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| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:day |
25
|
| pubmed:volume |
263
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
13486-9
|
| pubmed:dateRevised |
2009-11-19
|
| pubmed:meshHeading |
pubmed-meshheading:3417668-Adenosine Triphosphate,
pubmed-meshheading:3417668-Amino Acid Sequence,
pubmed-meshheading:3417668-Animals,
pubmed-meshheading:3417668-Binding Sites,
pubmed-meshheading:3417668-Brain,
pubmed-meshheading:3417668-Calcium,
pubmed-meshheading:3417668-Calmodulin,
pubmed-meshheading:3417668-Chromatography, Gel,
pubmed-meshheading:3417668-Chromatography, High Pressure Liquid,
pubmed-meshheading:3417668-Cyanogen Bromide,
pubmed-meshheading:3417668-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:3417668-Kinetics,
pubmed-meshheading:3417668-Molecular Sequence Data,
pubmed-meshheading:3417668-Peptide Fragments,
pubmed-meshheading:3417668-Phosphorylation,
pubmed-meshheading:3417668-Phosphoserine,
pubmed-meshheading:3417668-Phosphothreonine,
pubmed-meshheading:3417668-Protein Kinases,
pubmed-meshheading:3417668-Rats
|
| pubmed:year |
1988
|
| pubmed:articleTitle |
Ca2+/calmodulin-dependent protein kinase II. Identification of a regulatory autophosphorylation site adjacent to the inhibitory and calmodulin-binding domains.
|
| pubmed:affiliation |
Howard Hughes Medical Institute, Vanderbilt University School of Medicine, Nashville, Tennessee 37232.
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| pubmed:publicationType |
Journal Article
|