The selective utilization of substrates in vivo by the phosphatidylethanolamine and phosphatidylcholine biosynthetic enzymes Ept1p and Cpt1p in yeast

被引:29
作者
Boumann, HA
de Kruijff, B
Heck, AJR
de Kroon, AIPM
机构
[1] Ctr Biomembranes & Lipid Enzymol, Biomembrane Inst, Dept Membrane Biochem, NL-3584 CH Utrecht, Netherlands
[2] Bijvoet Ctr Biomol Res, Dept Biomol Mass Spectrometry, NL-3584 CA Utrecht, Netherlands
[3] Utrecht Inst Pharmaceut Sci, NL-3584 CA Utrecht, Netherlands
关键词
phosphatidylethanolamine; phosphatidylcholine; Kennedy pathway; diacylglycerol; on vivo deuterium pulse labeling; electrospray ionization tandem mass spectrometry;
D O I
10.1016/j.febslet.2004.05.043
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
In yeast, the aminoalcohol phosphotransferases Ept1p and Cpt1p catalyze the final steps in the CDP-ethanolamine and CDP-choline routes leading to phosphatidylethanolamine (PE) and phosphatidyleholine (PC), respectively. To determine how these enzymes contribute to the molecular species profiles of PE and PC in vivo, wild-type, cpt1Delta, and ept1Delta cells were pulse labeled with deuterated ethanolamine and choline. Analysis of newly synthesized PE and PC using electrospray ionization tandem mass spectrometry revealed that PE and PC produced by Ept1p and Cpt1p have different species compositions, demonstrating that the enzymes consume distinct sets of diacylglycerol species in vivo. Using the characteristic phospholipid species profiles produced by Ept1p and Cpt1p as molecular fingerprints, it was also shown that in vivo CDP-monomethylethanolarnine is preferentially used as substrate by Ept1p, whereas CDP-dimethylethanolamine and CDP-propanolamine are converted by Cpt1p. (C) 2004 Published by Elsevier B.V. on behalf of the Federation of European Biochemical Societies.
引用
收藏
页码:173 / 177
页数:5
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