The metabolism of 1‐acyl‐PAF in rabbit platelets and its possible interaction with PAF

AbstractThe metabolism of 1‐acyl‐2‐acetyl‐sn‐glycero‐3‐phosphocholine (1‐acyl‐PAF), a naturally occurring analogue of platelet activating factor (PAF), was investigated in rabbit platelets. Our studies showed that 1‐acyl‐[3H]PAF (1‐palmitoyl‐2‐acetyl‐sn‐glycero‐3‐phospho[N‐methyl‐3H]‐choline) was converted by platelets into phosphatidyl‐[3H]choline ([3H]PC) in a time‐dependent fashion. The formation of [3H]PC occurred at a rate similar to that observed when lyso‐[3H]PC (palmitoyl‐sn‐glycero‐3‐phospho[N‐methyl‐3H]choline) was used as substrate. In addition, a time‐dependent increase in the level of water‐soluble radioactivity was observed during the incubation of platelets with either 1‐acyl‐[3H]PAF or lyso‐[3H]PC. This increase was parallel to the formation of [3H]PC and was not observed in the presence of [14C]PAF (1‐octadecyl‐2‐acetyl‐sn‐glycerol‐3‐phospho[N methyl‐14C]choline). Analysis by thin‐layer chromatography showed that the soluble radioactivity was mainly associated with glycerophosphocholine (GPC). On the other hand, the preincubation of platelets with phenylmethylsulfonyl fluoride, an inhibitor of the acetylhydrolase, reduced the hydrolysis of 1‐acyl‐[3H]PAF to [3H]GPC with a concomitant accumulation of radioactivity in 1‐acyl‐PAF. These findings suggest that 1‐acyl‐PAF is converted into PC through deacetylation‐reacylation with lysoPC as an obligatory intermediate. The findings also indicate that the lysoPC resulting from 1‐acyl‐PAF is either reacylated to phosphatidylcholine (PC) or hydrolyzed to GPC by lysophospholipase. Finally, we showed that the stimulation of platelets with PAF led to a time‐ and concentration‐dependent increase in the conversion of 1‐acyl‐[3H]PAF to [3H]PC. The stimulatory effect of PAF was not observed when platelets were lysed before incubation, suggesting that PAF enhances the metabolism of 1‐acyl‐PAF, probably by accelerating its translocation through the plasma membrane.