title: Modulation of acetyl-CoA:1-alkyl-2-lyso-sn-glycero-3-phosphocholine (lyso-PAF) acetyltransferase in human polymorphonuclears. The role of cyclic AMP-dependent and phospholipid sensitive, calcium-dependent protein kinases.
abstract: In order to characterize the mechanism of activation of the enzyme 1-O-alkyl-2-lyso-sn-glycero-3-phosphocholine:acetyl-CoA acetyltransferase (EC 2.3.1.67) which is the limiting step in the regulation of the synthesis of the potent inflammatory mediator 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine; homogenates from human polymorphonuclear leukocytes were incubated in the presence of the catalytic subunit of cyclic AMP-dependent protein kinase and in the presence of a partially purified phospholipid sensitive, calcium-dependent protein kinase (PrKC). The first kinase was found to enhance up to 3-fold acetyltransferase activity in a dose- and time-dependent manner. In homogenates from PMN previously stimulated with complement-coated zymosan particles, the decay of acetyltransferase activity was partially prevented by the addition of soybean trypsin inhibitor and almost completely inhibited when the homogenates were supplemented with inhibitors of alkaline phosphatase such as 50 mM KF and 100 microM paranitrophenylphosphate. Under these conditions it was possible to initiate the decay of acetyltransferase activity by adding an excess of alkaline phosphatase. Preincubation of PMN with 12-O-tetradecanoylphorbol-13-acetate previous or simultaneously to the addition of ionophore A23187 reduced the increase in acetyltransferase produced by ionophore A23187, whereas the generation of superoxide anions was enhanced. Addition of partially purified PrKC to homogenates from ionophore A23187-stimulated PMN, reduced acetyltransferase activity by 63%, whereas only a 16% inhibition was observed on homogenates from resting PMN. These data indicate the modulation of acetyltransferase activity in human polymorphonuclear leukocytes by a phosphorylation-dephosphorylation mechanism linked to cyclic AMP-dependent protein kinase. Phospholipid sensitive, calcium-dependent protein kinase seems not to be involved in the mechanism of activation, but, most probably, in the generation of negative activation signals.
authors.affiliation: Instituto de Investigaciones Medicas de la Fundacion Jimenez Diaz.
authors.forename: M L
authors.initials: ML
authors.lastname: Nieto
authors.forename: S
authors.initials: S
authors.lastname: Velasco
authors.forename: M
authors.initials: M
authors.lastname: Sanchez Crespo
mesh_terms: D000123:Acetyltransferases / Q000097:blood* / Q000302:isolation & purification; D000469:Alkaline Phosphatase / Q000378:metabolism; D000001:Calcimycin / Q000494:pharmacology; D004789:Enzyme Activation; D006801:Humans; D007700:Kinetics; D009504:Neutrophils / Q000201:enzymology*; D011493:Protein Kinase C / Q000097:blood*; D011494:Protein Kinases / Q000097:blood*; D013481:Superoxides / Q000097:blood; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
chemical_list: D013481:Superoxides; D000001:Calcimycin; D000123:Acetyltransferases; C046035:1-alkylglycerophosphocholine acetyltransferase; D011494:Protein Kinases; D011493:Protein Kinase C; D000469:Alkaline Phosphatase; D013755:Tetradecanoylphorbol Acetate