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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
1
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pubmed:dateCreated |
1983-1-7
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pubmed:abstractText |
We developed a specific antibody to the catalytic subunit (C-subunit) of cyclic AMP-dependent protein kinase and used it to localize C-subunit in cultured cells. C-subunit antigen was purified from bovine cardiac muscle and cross-linked to hemocyanin with glutaraldehyde. Immunized goat serum showed a low titer of antibody after boosting; it was enriched 100-fold by affinity chromatography on catalytic subunit-Sepharose. The antibody immunoprecipitated C-subunit from type I and type II holoenzyme and depleted enzymatic activity from solution. At 12.5 nM antigen, 1 microgram antibody immunoprecipitated 10 ng of C-subunit. Immunoprecipitation of 35S-labeled cell extracts and 125I-antibody detection on nitrocellulose paper revealed that the antibody specifically reacts with C-subunit in Chinese hamster ovary (CHO) whole cell extracts. Using indirect immunofluorescence to localize C-subunit, we found a pattern of diffuse staining in the cytoplasm of CHO cells with little or no nuclear staining. A similar distribution of the enzyme was observed in Swiss 3T3 cells, bovine endothelial tracheal cells, human lung fibroblasts and NRK cells. Treatment of CHO cells with 8-bromo-cyclic AMP produced no change in the pattern or intensity of immunofluorescence. We conclude that the majority of C-subunit is localized in cytoplasm and that in cultured fibroblasts exposure to cyclic AMP analogues causes no apparent redistribution of catalytic subunit.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-1147218,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-1148240,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-164013,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-165513,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-16921,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-187175,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-189763,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-192719,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-196623,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-201274,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-205879,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-220219,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-222748,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-227897,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-27084,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-41953,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-4322607,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-4322609,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-4327009,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-4375761,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-4850204,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-4850305,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-6154700,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-6245082,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-6256373,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-6261262,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-6261328,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-6267050,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-6267595,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-6272695,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-6288732,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-7000793,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6754745-952867
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Oct
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pubmed:issn |
0021-9525
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
95
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
64-72
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pubmed:dateRevised |
2009-11-19
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pubmed:meshHeading |
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pubmed:year |
1982
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pubmed:articleTitle |
Localization of the catalytic subunit of cyclic AMP-dependent. Protein kinase in cultured cells using a specific antibody.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.
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