Synthesis of 1,2-diacyl-sn-glycerophosphatidylserine from egg phosphatidylcholine by phosphoramidite methodology.
A simple chemical method for the synthesis of 1,2-diacyl-sn-glycerophosphatidylserine (PS), with the same fatty acid composition in the sn-1 and sn-2 glycerol positions as egg phosphatidylcholine (PC), is described. PS synthesis was carried out by a phosphite-triester approach, using 2-cyanoethyl-N,N,N',N'-tetraisopropylphosphorodiamidite (phosphoramiditate) as the phosphorylating agent, for the formation of phosphate linkage between serine and diacylglycerol. 1,2-Diacylglycerol, obtained from PC hydrolysis by phospholipase C, was coupled with N-t-BOC-L-serinebenzhydryl ester phosphoramidite with tetrazole as catalyst. Phosphite-triester was oxidized to the corresponding phosphate-triester with 30% H2O2 in CH2Cl2. The cyanoethyl group was removed by addition of an Et3N/CH3 CN/pyridine mixture, and trifluoroacetic acid was used to eliminate the protecting groups of O-(1,2-diacylglycero-3-phospho)-N-t-BOC-serinebenzhydryl ester. Purified PS was identified by thin-layer chromatography, infrared, and 1H nuclear magnetic resonance.
Morillo
M
M
Department of Biochemistry and Physiology, Faculty of Chemistry, University of Barcelona, Spain.
Sagristá
L M
LM
Africa de Madariaga
M
M
Eritja
R
R
D002849:Chromatography, Gas / Q000379:methods; D002855:Chromatography, Thin Layer / Q000379:methods; D003016:Clostridium perfringens / Q000201:enzymology; D004530:Egg Yolk; D007202:Indicators and Reagents; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D015394:Molecular Structure; D009943:Organophosphorus Compounds / Q000633:toxicity; D010713:Phosphatidylcholines / Q000737:chemistry; D010718:Phosphatidylserines / Q000138:chemical synthesis* / Q000737:chemistry; D013055:Spectrophotometry, Infrared; D010738:Type C Phospholipases
D007202:Indicators and Reagents; D009943:Organophosphorus Compounds; D010713:Phosphatidylcholines; D010718:Phosphatidylserines; C049084:2-cyanoethyl-N,N,N',N'-tetraisopropylphosphorodiamidite; D010738:Type C Phospholipases