title: Substrate dependence of human neutrophil protein kinase C.
abstract: To determine the contribution of phosphate acceptor substrate to the pattern of activity of calcium-dependent, phospholipid-sensitive protein kinase (protein kinase C, PKC), we assayed cytosolic and particulate PKC activity for histone, troponin, myosin light chain (MLC), and endogenous cellular proteins in human neutrophils stimulated with phorbol myristate acetate (PMA), the chemotactic peptide n-formyl-methionyl-leucyl-phenylalanine (FMLP) and synergistic stimulation with both agonists. In general, phosphotransferase activity in neutrophil subfractions toward troponin and endogenous proteins paralleled that toward histone, but MLC was a poor substrate for PKC and the pattern of phosphotransferase activity differed from that seen with the other substrates. Furthermore, the phosphorylation of endogenous neutrophil cytosolic proteins increased significantly after stimulation with FMLP, suggesting an endogenous cytosolic substrate(s) which increased in concentration following stimulation. We conclude that histone is a useful phosphate acceptor for study of PKC activity in human neutrophils, but substrate variability occurs and may influence interpretation of results in assays of PKC activity.
authors.affiliation: Department of Pediatrics, Vanderbilt University Medical Center, Nashville, TN.
authors.forename: J C
authors.initials: JC
authors.lastname: Gay
authors.forename: E S
authors.initials: ES
authors.lastname: Stitt
mesh_terms: D006657:Histones / Q000378:metabolism; D006801:Humans; D009218:Myosins / Q000378:metabolism; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000201:enzymology*; D010766:Phosphorylation; D011493:Protein Kinase C / Q000097:blood*; D013379:Substrate Specificity; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014336:Troponin / Q000378:metabolism
chemical_list: D006657:Histones; D014336:Troponin; D009240:N-Formylmethionine Leucyl-Phenylalanine; D011493:Protein Kinase C; D009218:Myosins; D013755:Tetradecanoylphorbol Acetate