Abstract
Cytidine 5'-diphospho-1,2-diacyl-sn-glycerol (CDP-diacylglycerol; CDP-DG) is an important intermediate in the biosynthesis of the major glycerophosphate-based phospholipids of prokaryotes and eukaryotes. This compound is expensive to purchase and inefficient to prepare chemically. Radiolabeled CDP-diacylglycerol is unavailable commercially. We describe a simple and inexpensive method to synthesize [3H]CDP-DG enzymatically. The three-step enzymatic procedure includes phosphorylation of [3H]glycerol to sn-[3H]glycerol 3-phosphate (G3P) by glycerokinase,acylation of [3H]G3P to [3H]phosphatidic acid (PA) by G3P acyltransferase, and conversion of [3H]PA and CTP to [3H]CDP-DG by CDP-DG synthase. This procedure is considerably less labor intensive and less expensive than is chemical synthesis, and the yield is at least 30%.
Copyright 1998 Academic Press.
Publication types
-
Research Support, U.S. Gov't, P.H.S.
MeSH terms
-
Acylation
-
CDPdiacylglycerol-Serine O-Phosphatidyltransferase / metabolism
-
Cytidine Diphosphate Diglycerides / biosynthesis*
-
Cytidine Diphosphate Diglycerides / chemistry
-
Cytidine Triphosphate / metabolism
-
Escherichia coli / metabolism
-
Glycerol / metabolism
-
Glycerol Kinase / metabolism
-
Glycerol-3-Phosphate O-Acyltransferase / metabolism
-
Glycerophosphates / metabolism
-
Membrane Proteins*
-
Phosphatidic Acids / metabolism
-
Phosphorylation
-
Saccharomyces cerevisiae / metabolism
-
Transferases (Other Substituted Phosphate Groups) / metabolism
-
Tritium
Substances
-
Cytidine Diphosphate Diglycerides
-
Glycerophosphates
-
Membrane Proteins
-
Phosphatidic Acids
-
Tritium
-
Cytidine Triphosphate
-
alpha-glycerophosphoric acid
-
Glycerol-3-Phosphate O-Acyltransferase
-
Glycerol Kinase
-
Transferases (Other Substituted Phosphate Groups)
-
cardiolipin synthetase
-
CDPdiacylglycerol-Serine O-Phosphatidyltransferase
-
Glycerol