2n-fatty acids from phosphatidylcholine label sphingolipids-A novel role of phospholipase A2?

In order to find out whether there is a phospholipase A2 (PLA2)-mediated link between glycerophospholipids and sphingolipids, L929 cells were labeled with 1n-palmitoyl-2n-[1-14C]palmitoyl phosphatidylcholine for 16-18 h or 90 min. After labeling for 16-18 h, 14C-sphingomyelin (SM), 14C-ceramide and 14C-sphingosine were demonstrated on autoradiograms of thin layer chromatograms of untreated or mildly hydrolyzed lipid extracts in different chromatographic systems. Strong hydrolysis of labeled SM proved that both possible moieties of SM, sphingosine and acyl moiety, had been labeled. The identity of SM and its enzymatic degradation product, ceramide, was verified by mass spectrometry. The label in SM-derived ceramide was demonstrated on an autoradiogram after thin layer chromatography. The inhibitor of (dihydro)ceramide synthase fumonisin B1 suppressed the label in sphingolipids significantly during 16-18 h (ceramide and SM), as well as during 90-min labeling (SM). The presence of inhibitors of PLA2 (bromoenol lactone, aristolochic acid and quinacrine dihydrochloride) diminished the label in SM significantly during the 90-min labeling. These results demonstrate a close metabolic relationship between glycerophospholipids and sphingolipids and give evidence for a novel role of PLA2.