Enzymatic synthesis of [3H]Cytidine 5'-diphospho-1, 2-diacyl-sn-glycerol.
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%.
Zhao
M
M
Department of Biological Sciences, Wayne State University, Detroit, Michigan 48202, USA.
Rua
D
D
Hajra
A K
AK
Greenberg
M L
ML
D000215:Acylation; D010717:CDPdiacylglycerol-Serine O-Phosphatidyltransferase / Q000378:metabolism; D003567:Cytidine Diphosphate Diglycerides / Q000096:biosynthesis* / Q000737:chemistry; D003570:Cytidine Triphosphate / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D005990:Glycerol / Q000378:metabolism; D005991:Glycerol Kinase / Q000378:metabolism; D005992:Glycerol-3-Phosphate O-Acyltransferase / Q000378:metabolism; D005994:Glycerophosphates / Q000378:metabolism; D008565:Membrane Proteins; D010712:Phosphatidic Acids / Q000378:metabolism; D010766:Phosphorylation; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D017855:Transferases (Other Substituted Phosphate Groups) / Q000378:metabolism; D014316:Tritium
D003567:Cytidine Diphosphate Diglycerides; D005994:Glycerophosphates; D008565:Membrane Proteins; D010712:Phosphatidic Acids; D014316:Tritium; D003570:Cytidine Triphosphate; C029620:alpha-glycerophosphoric acid; D005992:Glycerol-3-Phosphate O-Acyltransferase; D005991:Glycerol Kinase; D017855:Transferases (Other Substituted Phosphate Groups); C100091:cardiolipin synthetase; D010717:CDPdiacylglycerol-Serine O-Phosphatidyltransferase; D005990:Glycerol