Comparison of the molecular species patterns of phosphatidic acid, CDP-diacylglycerols and phosphatidylinositol in potato tuber, pea leaf and soya-bean microsomes: Consequences for the selectivity of the enzymes catalyzing phosphatidylinositol biosynthesis

Abstract

Microsomes prepared from pea leaf, potato tuber or germinated soya-beans, were incubated for 30 min with [ 14 C] glycerol3-phosphate. In the three tissues, phosphatidic acid (PA), CDP-diacylglycerols (CMP-PA) and phosphatidylinositol (PI) were labelled and could be separated by TLC. After methylation of phosphatidic acid, or treatment of CMP-PA by a nucleotidase, the molecular species composition of the three lipid classes could be determined by radio-HPLC. The similarity observed between the distributions of radioactivity among CMP-PA and PA molecular species, in the three tissues, indicates that the enzyme CTP : PA cytidylyltransferase did not present any selectivity towards any molecular species of PA. In contrast, only two molecular species containing palmitic acid ( 16:0 18:2 and 16:0 18:3 ) were labelled in PI whereas labelled PA and CMP-PA contained molecular species possessing stearic acid ( 18:0 18:2 , 18:0 18:3 and 18:0 18:1 ). This indicates that the enzyme PI-synthase utilizes preferentially those molecular species of CMP-PA containing palmitic acid as substrates. However, mass analyses of PI prepared from the microsomes of the three tissues used in this study, indicated the presence of molecular species containing stearic acid ( 18:0 18:2 and 18:2 18:2 ). Except in soya-bean microsomes (where 18:0 18:2 -PI represented 16% of total PI), those last molecular species were always present in small amounts.