Incorporation and remodeling of extracellular phosphatidylcholine with short acyl residues in Saccharomyces cerevisiae

Abstract

The pem1/cho2 pem2/opi3 double mutant of Saccharomyces cerevisiae, which is auxotrophic for choline because of the deficiency in methylation activities of phosphatidylethanolamine, grew in the presence of 0.1 mM dioctanoyl-phosphatidylcholine (diC 8PC). Analysis of the metabolism of methyl- 13C-labeled diC 8PC (( methyl- 13C) 3-diC 8PC) by electrospray ionization tandem mass spectrometry (ESI-MS/MS) revealed that it was rapidly converted to ( methyl- 13C) 3-PCs containing C16 or C18 acyl chains. ( Methyl- 13C) 3-8:0-lyso-PC, ( methyl- 13C) 3-8:0-16:0-PC and ( methyl- 13C) 3-8:0-16:1-PC, which are the probable intermediate molecular species of acyl chain remodeling, appeared immediately after 5 min of pulse-labeling and decreased during the subsequent chase period. These results indicate that diC 8PC was taken up by the pem1 pem2 double mutant and that the acyl chains of diC 8PC were exchanged with longer yeast fatty acids. The temporary appearance of ( methyl- 13C) 3-8:0-lyso-PC suggests that the remodeling reaction may consist of deacylation and reacylation by phospholipase activities and acyltransferase activities, respectively. The detailed analyses of the structures of ( methyl- 13C) 3-8:0-16:0-PC and ( methyl- 13C) 3-8:0-16:1-PC by MS/MS and MS 3 strongly suggest that most ( methyl- 13C) 3-8:0-16:0-PCs have a C16:0 acyl chain at sn-1 position, whereas ( methyl- 13C) 3-8:0-16:1-PCs have a C16:1 acyl chain at either sn-1 or sn-2 position in a similar frequency, implying that the initial C16:0 acyl chain substitution prefers the sn-1 position; however, the C16:1 acyl chain substitution starts at both sn-1 and sn-2 positions. The current study provides a pivotal insight into the acyl chain remodeling of phospholipids in yeast.