Evidence for protein kinase C in bovine adrenocortical membrane preparations using (/sup 35/S) gamma-thio-ATP as a phosphate donor
Thio-substituted ATP is a sensitive probe for detecting protein kinase C activity as demonstrated in bovine adrenocortical cell membrane preparations. A single endogenous protein substrate with a molecular weight of approximately 47 Kd was rapidly phosphorylated with (/sup 35/S) gamma-thio-ATP as phosphate donor. Phosphorylation was significantly increased in 30 seconds and reached a plateau by 3 minutes. The activity of the endogenous membrane kinase was unaffected by ACTH, cAMP, calmodulin or trifluoperazine but was responsive to combinations of calcium (Ca), diolein and phosphatidyl serine (PS). In addition, the kinase was activated by the tumor promoting phorbol ester, 12-0-tetradecanoylphorbol-13-acetate, indicating that the membrane contains a protein kinase C and a single 47 Kd phosphorylatable protein substrate. The same substrate is phosphorylated by Ca/diolein/PS activated kinase in membrane preparations from a broad range of rat tissues. Attempts to identify the substrate indicate that it is neither the type I regulatory subunit of cAMP dependent protein kinase nor mitochondrial cytochrome P450.
- Research Organization:
- Wellesley College, MA
- OSTI ID:
- 5639614
- Journal Information:
- J. Cyclic Nucleotide Protein Phosphor. Res.; (United States), Journal Name: J. Cyclic Nucleotide Protein Phosphor. Res.; (United States) Vol. 11:6; ISSN JCNRE
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
560300 -- Chemicals Metabolism & Toxicology
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ANIMALS
ATP
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOLOGICAL EFFECTS
CARCINOGENS
CATTLE
CELL CONSTITUENTS
CELL MEMBRANES
CHEMICAL REACTIONS
DAYS LIVING RADIOISOTOPES
DOMESTIC ANIMALS
ENZYME ACTIVITY
ENZYMES
ESTERS
EVEN-ODD NUCLEI
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
MAMMALS
MEMBRANES
NUCLEI
NUCLEOTIDES
ORGANIC COMPOUNDS
PHORBOL ESTERS
PHOSPHORUS-GROUP TRANSFERASES
PHOSPHORYLATION
PHOSPHOTRANSFERASES
PROTEINS
RADIOISOTOPES
RESPONSE MODIFYING FACTORS
RUMINANTS
SUBSTRATES
SULFUR 35
SULFUR ISOTOPES
TRACER TECHNIQUES
TRANSFERASES
VERTEBRATES