Quantitation of protein kinase C by immunoblot-expression in different cell lines and response to phorbol esters

Journal Article · Thu Jan 01 00:00:00 EST 1987 · J. Cell. Physiol.; (United States)

DOI:https://doi.org/10.1002/jcp.1041300116· OSTI ID:6515544

Stabel, S; Rodriguez-Pena, A; Young, S; Rozengurt, E; Parker, P J

Antisera have been raised against human protein kinase C and also against a synthetic peptide based on the sequence of the bovine brain enzyme (LLNQEEGEYYNVPIPE). These antibodies react with protein kinase C from a number of species (human, murine, rat, rabbit, bovine), indicating substantial conservation of epitopes. These antisera have been used to quantitate directly protein kinase C by immunoblot analysis. The authors show here that there is a strict correlation between the levels of immunoreactive polypeptide and extractable calcium- and phospholipid-dependent kinase activity for various cell lines. Treatment of murine, rat, and human cells with phorbol dibutyrate was found to deplete levels of immunoreactive protein kinase C severely. A detailed study of the time course of this depletion in Swiss 3T3 cells shows that it follows precisely the loss of extractable activity. On exposure to 400 nM phorbol 12,13-dibutyrate protein kinase C was essentially undetectable by 40 hours; the half-life of this down-regulation was 6.7 hours. This data thus demonstrate that the loss of immunoreactive protein kinase C and of extractable calcium- and phospholipid-dependent kinase activity precisely parallels the phorbol ester induced down-regulation of binding and responsiveness in Swiss 3T3 cells.

• Research Organization: Imperial Cancer Research Fund, London, England
• OSTI ID: 6515544
• Journal Information: J. Cell. Physiol.; (United States), Vol. 130:1
• Country of Publication: United States
• Language: English

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BIOLOGICAL EFFECTS
PHOSPHOTRANSFERASES
RADIOASSAY
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FIBROBLASTS
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SULFUR 35
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CARBOXYLIC ACIDS
CARCINOGENS
CONNECTIVE TISSUE CELLS
DAYS LIVING RADIOISOTOPES
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ELECTRON CAPTURE RADIOISOTOPES
ENZYMES
ESTERS
EVEN-ODD NUCLEI
INTERMEDIATE MASS NUCLEI
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LIGHT NUCLEI
LIPOTROPIC FACTORS
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NUCLEI
ODD-EVEN NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
PHOSPHORUS-GROUP TRANSFERASES
PRIMATES
RADIOISOTOPES
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SOMATIC CELLS
SULFUR ISOTOPES
TRANSFERASES
VERTEBRATES
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