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Isozymic forms of rat brain CA/sup 2 +/-activated and phospholipid-dependent protein kinase

Conference · · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)
OSTI ID:6704677
;
Three forms of Ca/sup 2 +/-activated and phospholipid-dependent protein kinase (protein kinase C) were purified from the cytosolic fraction of rat brain. These enzymes, designated as type I, II, and III protein kinase C, all have the similar molecular weight of 80 Kd, bind (/sup 3/H)-phorbol dibutyrate in the presence of Ca/sup 2 +/, and undergo autophosphorylation in the presence of Ca/sup 2 +/, phosphatidylserine, and diolein. Autophosphorylation of these kinases resulted in an incorporation of 1- 1.5 mol /sup 32/P/mol of enzyme. Analysis of the /sup 32/P-labeled tryptic peptides derived from the autophosphorylated protein kinase C by two-dimensional peptide mapping revealed that these kinases had different sites of autophosphorylation. Phosphoamino acid analysis revealed that the type I and type III protein kinase C mainly phosphorylated at Ser residue while the type II kinase phosphorylated at both Ser and Thr residues. In addition, polyclonal antibodies previously prepared against a mixed enzyme fraction preferentially inhibited the type I and type II enzymes but less effectively toward the type III enzyme. Monoclonal antibody specifically against the type II protein kinase C did not inhibit the type I or type III enzymes. These kinases also had different susceptibility to limited proteolysis by trypsin and upon proteolytic degradation they generate distinct fragments. These results demonstrate the presence of isozymic forms of protein kinase C in rat brain.
Research Organization:
National Institutes of Health, Bethesda, MD
OSTI ID:
6704677
Report Number(s):
CONF-8606151-
Conference Information:
Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Journal Volume: 45:6
Country of Publication:
United States
Language:
English

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Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ALKALINE EARTH METAL COMPOUNDS
AMINO ACIDS
ANIMALS
ANTIBODIES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BODY
BRAIN
CALCIUM COMPOUNDS
CARBOXYLIC ACIDS
CARCINOGENS
CATIONS
CENTRAL NERVOUS SYSTEM
CHARGED PARTICLES
CHEMICAL REACTIONS
DAYS LIVING RADIOISOTOPES
DECOMPOSITION
ENZYMES
ESTERS
IONS
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LABELLED COMPOUNDS
LIGHT NUCLEI
LIPIDS
MAMMALS
MOLECULAR WEIGHT
MONOCLONAL ANTIBODIES
NERVOUS SYSTEM
NUCLEI
ODD-ODD NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC PHOSPHORUS COMPOUNDS
ORGANS
PHORBOL ESTERS
PHOSPHOLIPIDS
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PHOSPHORUS-GROUP TRANSFERASES
PHOSPHORYLATION
PHOSPHOTRANSFERASES
PROTEOLYSIS
PURIFICATION
RADIOISOTOPES
RATS
REACTION KINETICS
RODENTS
TRACER TECHNIQUES
TRANSFERASES
TRITIUM COMPOUNDS
VERTEBRATES