The yeast phospholipid N-methyltransferases catalyzing the synthesis of phosphatidylcholine preferentially convert di-C16:1 substrates both in vivo and in vitro.
Phosphatidylcholine (PC) is an important and abundant structural component of the membranes of eukaryotic cells. In the yeast Saccharomyces cerevisiae, the primary route for the biosynthesis of PC consists of three consecutive methylation steps of phosphatidylethanolamine (PE) catalyzed by the phospholipid N-methyltransferases Cho2p and Opi3p. To investigate how these biosynthetic enzymes contribute to the composition of the PC species profile, the precursor-product relationships between PE and newly synthesized PC were determined at the level of the molecular species by using electrospray ionization tandem mass spectrometry and stable isotope labeling. In vivo labeling of yeast cells for 10 min with [methyl-D3]methionine revealed the preferential methylation of di-C16:1 PE over a range of PE species compositions. A similar preferential conversion of di-C16:1 PE to PC was found in vitro upon incubating isolated microsomes with S-adenosyl[methyl-D3]methionine. Yeast opi3 and cho2 deletion strains were used to distinguish between the substrate selectivities of Cho2p and Opi3p, respectively. Both biosynthetic enzymes were found to participate in the speciesselective methylation with Cho2p contributing the most. The combined results indicate that the selective methylation of PE species by the methyltransferases plays an important role in shaping the steady-state profile of PC molecular species in yeast.
Boumann
Henry A
HA
Department Biochemistry of Membranes, Centre for Biomembranes and Lipid Enzymology, Institute of Biomembranes, Padualaan 8, 3584 CH Utrecht, The Netherlands. h.a.boumann@chem.unn.nl
Chin
Patrick T K
PT
Heck
Albert J R
AJ
De Kruijff
Ben
B
De Kroon
Anton I P M
AI
D001669:Biochemical Phenomena; D001671:Biochemistry; D002384:Catalysis; D017353:Gene Deletion; D066298:In Vitro Techniques; D008745:Methylation; D008780:Methyltransferases / Q000737:chemistry / Q000502:physiology*; D008954:Models, Biological; D050941:Phosphatidyl-N-Methylethanolamine N-Methyltransferase; D010713:Phosphatidylcholines / Q000737:chemistry*; D010714:Phosphatidylethanolamines / Q000737:chemistry; D010743:Phospholipids / Q000378:metabolism; D012436:S-Adenosylmethionine / Q000737:chemistry; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D021241:Spectrometry, Mass, Electrospray Ionization; D013379:Substrate Specificity; D013997:Time Factors
D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D010743:Phospholipids; C483858:phosphatidylethanolamine; D012436:S-Adenosylmethionine; D008780:Methyltransferases; D050941:Phosphatidyl-N-Methylethanolamine N-Methyltransferase