title: Biosynthesis of paf-acether. IX. Role for a phosphorylation-dependent activation of acetyltransferase in antigen-stimulated mouse mast cells.
abstract: Mouse bone marrow-derived mast cells passively sensitized with monoclonal IgE released paf-acether (platelet-activating factor) and beta-hexosaminidase when challenged with the specific antigen. The formation and the release of paf-acether followed an early increase in the activity of the acetyltransferase, the main enzyme in paf-acether biosynthesis. The antigen-induced activation of the acetyltransferase was dependent on physiologic temperature and on the presence of Ca2+. By using microsomal fractions from unchallenged and challenged mast cells, the Vmax values were 3.5 and 12.0 nmol/min/mg of protein, respectively, whereas in both cases a Km value for acetyl-coenzyme A of 172 microM was measured. The stimulation of acetyltransferase could be mimicked in vitro under experimental conditions which favor phosphorylation, i.e. adding ATP and Mg2+ to lysates from unchallenged mast cells. In contrast, ATP and Mg2+ were uneffective on lysates from challenged cells that exhibited high level of acetyltransferase activity, suggesting that phosphorylation of the enzyme already took place at the time of cell stimulation. Moreover, addition of alkaline phosphatase to microsomal fraction obtained from either antigen-challenged mouse bone marrow-derived mast cells or unchallenged cells, resulted in 52% and 43% loss of acetyltransferase activity, respectively. Phorbol myristate acetate treatment of cells doubled the enzyme activity supporting the phosphorylation hypothesis. Thus, we report on the immunologic activation of a key enzyme for paf-acether synthesis and on the mechanism of this activation in a pure mast cell population. A link between bridging of IgE receptors and the activation of an enzyme critical to the formation of a lipid mediator is thereby evidenced.
mesh_terms: D000123:Acetyltransferases / Q000378:metabolism*; D000818:Animals; D000941:Antigens / Q000276:immunology; D001854:Bone Marrow Cells; D002118:Calcium / Q000494:pharmacology; D002460:Cell Line; D004789:Enzyme Activation; D007073:Immunoglobulin E / Q000502:physiology; D007700:Kinetics; D008407:Mast Cells / Q000502:physiology*; D051379:Mice; D008861:Microsomes / Q000201:enzymology; D010766:Phosphorylation; D010972:Platelet Activating Factor / Q000096:biosynthesis*; D013347:Subcellular Fractions / Q000201:enzymology
chemical_list: D000941:Antigens; D010972:Platelet Activating Factor; D007073:Immunoglobulin E; D000123:Acetyltransferases; C046035:1-alkylglycerophosphocholine acetyltransferase; D002118:Calcium