Rapid accumulation of diacyl lipid in rat liver microsomes by selective acylation of 2-acyl-sn-glycero-3-phosphorylserine

Abstract

2-Acyl- sn-glycero-3-phosphoryl[U- 14C]serine (2-acylGPS) was prepared by digesting 1,2-dioleoyl-sn-glycero-3-phosphoryl[U- 14C]serine, mixed with pig brain phosphatidylserine (PS), with Rhizopus arrhizus lipase. The labeled lysolipid was shown to be actively acylated by rat liver microsomes in the presence of acyl-CoA thioesters, with the formation of diacylPS. In experiments with unlabeled lysolipid and [ 14C]acyl-CoA thioesters, digestion of the products with pancreatic phospholipase A 2 showed that the acyl groups had been incorporated almost exclusively at position-1. The acylation reactions were dependent on pH of the incubation medium, time of incubation, concentration of microsomal protein and of lysolipid and thioester substrates. With [ 14C]lysolipid there was a clear preference for long-chain saturated over unsaturated acyl group donors and highest acylation rates were recorded with stearoyl-CoA. Since stearate is the major fatty acid at position-1 of tissue PS, these findings suggest that deacylation-reacylation reactions may be of importance in regulating the fatty acid profile of this lipid. The observed rates of acylation of 2-acylGPS in vitro were high in relation to the endogenous level of PS in the microsomal membranes, and short-term incubations led to net synthesis of PS with a many-fold increase in its concentration. The question is raised whether this is a latent activity expressed under optimal assay conditions in vitro, conditions that may not obtain in a cell, or whether the high acylating potential is indicative of an important function for the addition, or turnover, of acyl groups at position-1 of the serine lipid.