J. Lipid Res.
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Journal of Lipid Research, Vol 21, 1053-1057, Copyright © 1980 by Lipid Research, Inc.


ARTICLES

An improved procedure for the synthesis of 14C-labeled phosphatidylserine from cerebral phosphatidic acid

P Orlando, G Ippolito, L Binaglia, C Giordano and G Porcellati

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.
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