Partial purification and characterization of the calcium-dependent and phospholipid-dependent protein kinase C from chick oviduct.
The calcium-dependent and phospholipid-dependent protein kinase C is partially purified (2000-fold) from chick oviduct cytosol by chromatography on DEAE-cellulose, Sephadex G150 and (after affinity binding of the enzyme to phosphatidylserine-diolein liposomes) on Bio-Gel A-0.5m. The enzyme is activated by phosphatidylserine and calcium ions. Diolein increases the affinity of the enzyme for both cofactors and can be replaced by the tumour-promoting phorbol ester 12-O-tetradecanoyl-phorbol 13-acetate. Phosphatidylserine can be replaced by unsaturated long-chain fatty acids. Quercetin and phloretin inhibit the kinase reversibly competing with ATP. Several oviduct proteins are found to be phosphorylated by the partially purified enzyme. One of the substrates, a 78-kDa protein, appears to be ovotransferrin.
Horn
F
F
Gschwendt
M
M
Marks
F
F
D000818:Animals; D001665:Binding Sites / Q000187:drug effects; D001667:Binding, Competitive; D002118:Calcium / Q000494:pharmacology*; D002645:Chickens; D002848:Chromatography, DEAE-Cellulose; D003600:Cytosol / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D010057:Oviducts / Q000201:enzymology*; D010743:Phospholipids / Q000494:pharmacology*; D011493:Protein Kinase C; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases / Q000302:isolation & purification*; D013379:Substrate Specificity
D010743:Phospholipids; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases; D011493:Protein Kinase C; D002118:Calcium