An improved procedure for the synthesis of 14C-labeled phosphatidylserine from cerebral phosphatidic acid.
A complete procedure to prepare a highly labeled phosphatidyl-L-[U-14C]serine possessing the same fatty acid composition of brain phospholipids is reported. CDP-diglyceride was synthesized by reaction between phosphatidic acid and CMP-morpholidate as the dicyclohexylcarboxamidium salt. The reaction between CDP-diglyceride and L-[U-14C]serine to produce the labeled phosphatidylserine was catalyzed by the CDP-diglyceride: L-serine phosphatidyl transferase (EC 2.7.8.8) from E. coli. A selective inhibition of phosphatidylserine decarboxylase activity, present as contaminant in the enzyme extract, was introduced in order to avoid a low yield of product. Traces of phosphatidylethanolamine (about 1%) were easily removed by preparative thin-layer chromatography. The yield of the labeled product was as high as 87% and it specific radioactivity was 170 mCi/mmol.
D000818:Animals; D010717:CDPdiacylglycerol-Serine O-Phosphatidyltransferase; D002262:Carboxy-Lyases / Q000037:antagonists & inhibitors; D002417:Cattle; D055598:Chemical Phenomena; D002621:Chemistry; D002855:Chromatography, Thin Layer; D003567:Cytidine Diphosphate Diglycerides; D004926:Escherichia coli / Q000201:enzymology; D010712:Phosphatidic Acids; D010718:Phosphatidylserines / Q000037:antagonists & inhibitors / Q000138:chemical synthesis*; D010770:Phosphotransferases
D003567:Cytidine Diphosphate Diglycerides; D010712:Phosphatidic Acids; D010718:Phosphatidylserines; D010770:Phosphotransferases; D010717:CDPdiacylglycerol-Serine O-Phosphatidyltransferase; D002262:Carboxy-Lyases; C022353:phosphatidylserine decarboxylase