ENZYMATIC-SYNTHESIS OF [H-3]CYTIDINE 5'-DIPHOSPHO-1,2-DIACYL-SN-GLYCEROL

Cytidine 5'-diphospho-1,2-diacyl-sn-glycerol (CDP-diacylglycerol; CDP- DQ is an important intermediate in the biosynthesis of the major glyce rophosphate-based phospholipids of prokaryotes and eukaryotes, This co mpound is expensive to purchase and inefficient to prepare chemically. Radiolabeled CDP-diacylglycerol is unavailable commercially. We descr ibe a simple and inexpensive method to synthesize [H-3]CDP-DG; enzymat ically, The three step enzymatic procedure includes phosphorylation of [H-3]glycerol to sn-[H-3]glycerol 3-phosphate (G3P) by glycerokinase, acylation of [H-3]G3P to [H-3]phosphatidic acid (PA) by G3P acyltrans ferase, and conversion of [H-3]PA and CTP to [H-3]CDP-DG by CDP-DG syn thase. This procedure is considerably less labor intensive and less ex pensive than is chemical synthesis, and the yield is at least 30%. (C) 1998 Academic Press.