linoleic-acid and lysophosphatidylethanolamine

Evidence was obtained for a CoA-dependent transfer of linoleate from rat lung microsomal phosphatidylcholine to lysophosphatidylethanolamine without the intervention of a Ca2+-requiring phospholipase A2 activity and ATP. To study this CoA-mediated transacylation process, microsomes were prepared in which the endogenous phosphatidylcholine was labeled by protein-catalyzed exchange with phosphatidylcholines containing labeled fatty acids in the sn-2-position. The apparent Km for CoA in the transfer of arachidonate from phosphatidylcholine to 1-acyllysophosphatidylethanolamine was 1.5 microM. At saturating lysophosphatidylethanolamine concentrations, the transacylation was linear with the amount of microsomal protein, i.e., a fixed percentage of the labeled fatty acid was transferred independent of the amount of microsomal protein. A maximal transfer of 12.2% for arachidonate and 2.0% for linoleate from the respective phosphatidylcholines to lysophosphatidylethanolamine was observed in 30 min. With 1-acyl-2-[1-14C]arachidonoylphosphatidylcholine as acyl donor, lysophosphatidylethanolamine was the best acceptor followed by lysophosphatidylglycerol and lysophosphatidylserine. Lysophosphatidate barely functioned as acceptor. These data provide further evidence for the widespread occurrence of CoA-mediated transacylation reactions. The arachidonate transacylation from phosphatidylcholine to other phospholipids in lung tissue may contribute to the low level of arachidonate in pulmonary phosphatidylcholine.

Topics: Acylation; Animals; Arachidonic Acid; Arachidonic Acids; Coenzyme A; Fatty Acids; In Vitro Techniques; Linoleic Acid; Linoleic Acids; Lung; Lysophospholipids; Male; Microsomes; Phosphatidylcholines; Phosphatidylethanolamines; Rats; Rats, Inbred Strains; Time Factors

When 600 X g supernatants of 10% (w/v) rat lung homogenates were incubated with CDP[Me-14C]choline both saturated and unsaturated species of phosphatidylcholine were formed from endogenous diacylglycerols. The percentage radioactivity in the disaturated species of total phosphatidylcholine increased with time from 12% after 5 min to 30% after 60 min incubation. In similar experiments with 20 000 X g supernatants, the increase in the disaturated species of microsomal phosphatidylcholine was from 25 to 37% over the same time period. In incubations of isolated microsomes in buffer, the percent of 14C label in disaturated phosphatidylcholine remained constant at a level of 25%. To investigate a possible role of cytosolic factor(s) in the increase in the percentage of disaturated phosphatidylcholine with time, microsomes were prelabeled by incubation in buffer with CDP[Me-14C]choline to give a fixed ratio of radioactive saturated and unsaturated phosphatidylcholine species. When the reisolated microsomes were incubated in buffer, the distribution of radioactivity over saturated and unsaturated species remained constant. In contrast, incubation of prelabeled microsomes in the presence of cytosol caused an increase in the percent radioactivity in saturated phosphatidylcholines from a starting value of 18 to 30% after 60 min incubation, while leaving total phosphatidylcholine radioactivity unaffected. These results indicate a remodeling of phosphatidylcholine under the influence of a cytosolic factor(s). Evidence is presented that suggests that Ca2+-independent-cytosolic phospholipase A2 activity as well as a microsomal ATP-independent CoA-mediated acyltransferase activity might contribute to this remodeling. The cytosol donates the necessary CoA for this acyl transfer as well as saturated acyl-CoA for the reacylation of lysophosphatidylcholine.

Topics: Acyl Coenzyme A; Animals; Coenzyme A; Cytosol; In Vitro Techniques; Linoleic Acid; Linoleic Acids; Lung; Lysophospholipids; Male; Microsomes; Phosphatidylcholines; Phosphatidylethanolamines; Phospholipids; Rats; Rats, Inbred Strains