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

Conference · Thu May 01 00:00:00 EDT 1986 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)

OSTI ID:6704677

Huang, K P; Huang, F L

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-
• Journal Information: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 45:6; Conference: 76. annual meeting of the Federation of American Society for Experimental Biology, Washington, DC, USA, 8 Jun 1986
• Country of Publication: United States
• Language: English

Related Subjects

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