title: Studies on protein kinase C tightly-bound to rat liver plasma membrane and its protease-activated form.
abstract: 1. Rat liver plasma membrane contained two types of protein kinase C which could be extracted by Ca2(+)-chelator and detergent, respectively. The activities of these two enzymes were nearly equivalent. 2. The detergent-extracted protein kinase C, tightly-bound to membrane, was separated into two subtypes by hydroxyapatite column chromatography. Based on the elution profile and the Ca2+/phospholipid requirement, the major and the minor components were identified as type III and type II protein kinase C, respectively. 3. The detergent-extracted protein kinase C was converted to an active fragment with Mr 45,000 by limited proteolysis with trypsin. Incubation under physiological level of ionic strength increased the stability of this active enzyme and protected it from further inactivation by trypsin. 4. Phosphorylation of H1 histone by the protease-activated kinase was stimulated 1.5-2-fold by phosphatidylserine. However, this enzyme phosphorylated multiple proteins in rat liver subcellular fractions in Ca2(+)- and phospholipid-independent manner. 5. These results suggest that the protein kinase C (mainly type III enzyme) tightly-bound to rat liver plasma membrane may have important role through protein phosphorylation by the native or the protease-activated kinase.
authors.affiliation: Department of Biochemistry, Fukui Medical School, Japan.
authors.forename: E
authors.initials: E
authors.lastname: Hashimoto
authors.forename: F
authors.initials: F
authors.lastname: Takeuchi
authors.forename: H
authors.initials: H
authors.lastname: Yamamura
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000201:enzymology; D010450:Endopeptidases / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D066298:In Vitro Techniques; D008099:Liver / Q000201:enzymology*; D008970:Molecular Weight; D011493:Protein Kinase C / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains
chemical_list: D011493:Protein Kinase C; D010450:Endopeptidases