Isolation and partial characterization of the neutral glycosphingolipids and gangliosides of the human heart

Levis, Gabriel M.; Karli, J; Moulopoulos, S.D.

doi:10.1007/bf02533559

Cited by 14 publications

(1 citation statement)

References 33 publications

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“…It must be noted that a small amount of residual I13NeuAc-LacCer was also detected. As shown before [38], the thin-layer system used could not differentiate between various gangliosides, therefore further work is needed for the characterization of the complex gangliosides.…”

Section: Incorporation O J [ U-14c]galactose Und [ U-'4c]glucosamine mentioning

confidence: 99%

Studies on Glycosphingolipid Biosynthesis by Lectin-Stimulated Human Lymphocytes

et al. 1980

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1. Human peripheral lymphocytes, normal and stimulated by phytohemagglutinin or concanavalin A, were investigated with respect to their ability to biosynthesize neutral and acidic glycosphingolipids from D‐[U‐14C]glucose, D‐[U‐14C]galactose and D‐[U14C]glucosamine as precursors. 2. Galactose and glucosamie are taken up selectively, in the presence of excessive glucose concentrations. Labeling of total neutral glycosphingolipids from D‐[U‐14C]galactose in normal cells decreases after the first 6 h while in stimulated cells there is a fourfold and a sevenfold increase after 6 and 12 h of incubation respectively. Under similar conditions stimulation with concanavalin A gives a fourfold increase after 12 h of incubation. 3. Analysis of individual glycosphingolipids biosynthesized from [U‐14C]galactose shows that lactosylceramide is the major radioactive neutral glycosphingolipid and that stimulation by phytohemagglutinin yields an almost sevenfold and 13‐fold increase in the radioactivity incorporated within 6 and 12 h of incubation respectively. Glucosylceramide shows about an eightfold increase, globotetraosylceramide a threefold increase and globotriaosylceramide a fourfold increase. The rate of incorporation into glucosylceramide of stimulated lymphocytes declines after 6 h of incubation, accompanied by a concomitant increase of incorporation into lactosylceramide. 4. At 12 h of incubation the ratios of radioactivities incorporated into neutral glycosphingolipids of phytohemagglutinin‐stimulated cells compared to normal cells were 1.0 for D‐[U‐14C]glucose 7.0 for D‐[U‐14C]galactose and 2.5 for D‐[U‐14C]glucosamine. Respective ratios for lactosylceramide are 1.0 for [U‐14C]glucose and 13.0 for [U‐14C]galactose. These differences did not arise from changes of the uptake of the glycosyl precursor by the cell due to stimulation. 5. Incorporation of D‐[U‐14C]galactose into II3‐N‐acetylneuraminosyllactosylceramide by cells stimulated by phytohemagglutinin and concanavalin A is enhanced 14‐times and 15‐times respectively. With [U‐14C]glucosamine as precursor, this increase in the labeling is much more impressive, 80‐fold after 12 h of incubation by the phytohemagglutinin‐stimulated lymphocytes. Neuraminidase treatment and gas radiochromatographic analysis of the labeled compound derived from [U‐14C]‐galactose as precursor indicated that 89% of the radioactivity was incorporated into the glucosyl and galactosyl moieties, in a ratio 1:1. With [U‐14C]glucosamine as precursor, a selective labeling of the sialyl moiety of the II3‐N‐acetylneuraminosyllactosylceramide was indicated. 6. The pattern of complex gangliosides (more complex than II3‐N‐acetylneuraminosyllactosylceramide) which are biosynthesized after phytohemagglutinin stimulation of the cells, show no significant differences when compared to the patterns obtained from normal cells.

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Section: Incorporation O J [ U-14c]galactose Und [ U-'4c]glucosamine mentioning

confidence: 99%

Studies on Glycosphingolipid Biosynthesis by Lectin-Stimulated Human Lymphocytes

et al. 1980

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Expression of neutral glycosphingolipids and gangliosides in human skeletal and heart muscle determined by indirect immunofluorescence staining

et al. 1994

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The expression of neutral glycosphingolipids and gangliosides has been studied in human skeletal and heart muscle using indirect immunofluorescence microscopy. Transversal and longitudinal cryosections were immunostained with specific monoclonal and polyclonal antibodies against the neural glycosphingolipids lactosylceramide, globoside, Forssman glycosphingolipid, gangliotetraosylceramide, lacto-N-neotetraosylceramide and against the gangliosides GM3(Neu5Ac) and GM1(Neu5Ac). To confirm the lipid nature of positive staining, control sections were treated with methanol and chloroform:methanol (1:1) before immunostaining. These controls were found to be either negative or strongly reduced in fluorescence intensity, suggesting that lipid bound oligosaccharides were detected. In human skeletal muscle, lactosylceramide was found to be the main neutral glycosphingolipid. Globoside was moderately expressed, lacto-N-neotetraosylceramide and gangliotetraosylceramide were minimally expressed and Forssman glycosphingolipid was not detected in human skeletal muscle. The intensities of the immunohistological stains of GM3 and GM1 correlated to the fact that GM3 is the major ganglioside in skeletal muscle whereas GM1 is expressed only weakly. In human heart muscle globoside was the major neutral glycosphingolipid. Lactosylceramide and lacto-N-neotetraosylceramide were moderately expressed, gangliotetraosylceramide was weakly expressed and the Forssman glycosphingolipid was not expressed at all in cardiac muscle. GM3 and GM1 were detected with almost identical intensity. All glycosphingolipids were present in plasma membranes as well as at the intracellular level.

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Gangliosides in membranes fromTorpedo electric organ

Hann

Evans

McCluer

et al. 1996

Lipids

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The electric organ membrane has been the subject of many studies, due principally to its rich content of nicotinic acetylcholine receptor (AChR). Knowing its lipid composition is clearly important. Although its major membrane lipids have been characterized, its ganglioside composition has not been as well-described. In this study, gangliosides were characterized in membranes prepared from two species of electric organ, Torpedo californica and T. nobiliana. The ganglioside content of total electric organ membranes and AChR-enriched membranes was similar in both species, accounting for from 0.9 to 1.5% of membrane lipid by weight. However, the AChR-enriched membranes contained significantly less ganglioside relative to AChR than did the total membrane preparations. Five major gangliosides were purified from T. californica and identified as II3NeuNAc-GgOse3 (GM2); II3(NeuNAc)2-GgOse3 (GD2), IV3NeuNAc, II3NeuNAc-GgOse4 (GD1a), IV3NeuNAc, II3(NeuNAc)2-GgOse4 (GT1b), and IV3(NeuNAc)2,II3(NeuNAc)2-GgOse4 (GQ1b). Together these five gangliosides accounted for over 90% of the total ganglioside present in the two membrane preparations from both species. The most abundant ganglioside by far was GM2, which accounted for about one-half of the ganglioside content, followed by GD2. Determination of the N-fatty acid composition was performed on gangliosides purified from T. nobiliana. The lower-order gangliosides, GM2, GD2, and GD1a, contained substantial amounts of very long chain fatty acids (> 20 carbons), including alpha-hydroxynervonic acid (15-21% of total). In contrast, unsubstituted, 14-18 carbon chains accounted for about 90% of the fatty acids on the two higher-order gangliosides, GT1b and GQ1b.

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Isolation and partial characterization of the neutral glycosphingolipids and gangliosides of the human heart

Cited by 14 publications

References 33 publications

Studies on Glycosphingolipid Biosynthesis by Lectin-Stimulated Human Lymphocytes

Studies on Glycosphingolipid Biosynthesis by Lectin-Stimulated Human Lymphocytes

Expression of neutral glycosphingolipids and gangliosides in human skeletal and heart muscle determined by indirect immunofluorescence staining

Gangliosides in membranes fromTorpedo electric organ

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