Lipid Composition of PC 12 Pheochromocytoma Cells: Characterization of Globoside as a Major Neutral Glycolipid

Toshio Ariga, Lawrence J. Macala, Megumi Saito, Renee K. Margolis, Lloyd A. Greene, Richard U. Margolis, Robert K Yu

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Abstract

We have studied the lipid composition of PC 12 pheochromocytoma cells cultured in the presence and absence of nerve growth factor (NGF). Neutral and acidic lipid fractions were isolated by column chromatography on DEAE-Sephadex and analyzed by high-performance thin-layer chromatography (HPTLC). The total lipid concentration was approximately 220 jug/mg of protein, and the concentration of neutral glycolipids was 1.6-1.8 Mg/mg of protein for both NGF-treated and untreated cells. The neutral glycolipid fraction contained a major component, which accounted for approximately 80% of the total and which was characterized as globoside on the basis of HPTLC mobility, carbohydrate analysis, fast atom bombardment mass spectrometry, and mild acid hydrolysis. The major fatty acids of globoside were Cl6:0 (10%), C18:0 (16%), C22:0 (23%), C24:l (17%), and C24:0 (24%). C18 sphingenine accounted for almost all of the long-chain bases. The other neutral glycolipids were tentatively identified as glucosylceramide (15%), lactosylceramide (4%), and globotriosylceramide (4.5%). The concentration of ganglioside sialic acid was approximately 0.34 and 0.18 p%f mg of protein for cells grown in the presence and absence of NGF, respectively. Although there was an increase in ganglioside concentration in NGF-treated cells, NGF did not produce any differential effects on the relative proportions of the individual gangliosides. Several of the gangliosides appear to contain fucose, and one of these was tentatively identified as fucosyl-GMltBrain-type gangliosides of the ganglio series were also detected by an HPTLC-immunostaining method. However, the fatty acid and long chain base compositions of PC 12 cell gangliosides (and their TLC mobility) differ from those of brain gangliosides. The major lipids of PC 12 cells, which were not significantly affected by NGF treatment, were phosphatidylcholine (40%), phosphatidylethanolamine (22%), and cholesterol (13%).

Original languageEnglish (US)
Pages (from-to)52-58
Number of pages7
JournalBiochemistry
Volume27
Issue number1
DOIs
Publication statusPublished - Jan 1 1988
Externally publishedYes

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ASJC Scopus subject areas

  • Biochemistry

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