Ca2+-dependent and Ca2+-independent pathways for release of arachidonic acid from phosphatidylinositol in endothelial cells.
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DOI: 10.1016/S0021-9258(18)45171-X
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摘要: The pathways for degradation of phosphatidylinositol (PI) were investigated in sonicated suspensions prepared from confluent cultures bovine pulmonary artery endothelial cells. time courses formation 3H-labeled and 14C-labeled metabolites phosphatidyl-[3H]inositol ([3H]Ins-PI) 1-stearoyl-2-[14C] arachidonoyl-PI determined at 37 degrees C pH 7.5 the presence 2 mM EDTA with or without a excess Ca2+. rates lysophosphatidyl-[3H]inositol ([3H]Ins-lyso-PI) 1-lyso-2-[14C] similar absence Ca2+, absolute amounts two radiolabeled lyso-PI products formed nearly identical. This indicated that was by phospholipase A1, A2 not measurable. In EDTA, [14C]arachidonic acid release 1-stearoyl-2-[14C]arachidonoyl-PI paralleled glycerophospho-[3H]inositol ([3H]GPI) [3H]Ins-PI. Formation [3H]GPI inhibited treatment specific sulfhydryl reagent, 2,2'-dithiodipyridine, this accompanied an increase [3H]Ins-lyso-PI. [14C] arachidonic increased 2-fold associated Ca2+-dependent activity. Under these conditions, [3H]inositol monophosphate production exceeded acid-labeled products, diacylglycerol plus monoacylglycerol, amount equal to [3H]GPI. Low concentrations phenylmethanesulfonyl fluoride (15-125 microM) release, decrease matched equivalent 14C label monoacyclglycerol. These data supported existence PI cells; A1-lysophospholipase pathway Ca2+-independent C-diacylglycerol lipase Ca2+-dependent. mean percentage released via Ca2+ 65 +/- 8%. nonpolar metabolized free 28 3%.
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