Isolation and structural characterization of glycosphingolipids of in vitro propagated bovine aortic endothelial cells

Abstract

Neutral glycosphingolipids and gangliosides were isolated from 3.7 x 10 9 primary bovine aortic endothelial cells and structurally characterized by immunological and chemical methods. Glucosyl-and lactosylceramide were detected as the main neutral glycosphingolipids (28% and 40% of total orcinol stain, respectively); LcOse 3 Cer and nLcOse 4 Cer were expressed to somewhat minor amounts (16% and 10% of total orcinol stain, respectively), and nLcOse 6 Cer occurred only in trace quantities. No neutral glycosphingolipids of the ganglio-series (GgOse 3 Cer and GgOse 4 Cer) and the globo-series (GbOse 4 Cer and the Forssman antigen) have been detected; only traces of GbOse 3 Cer were identified by TLC immunostaining. Positive CD15 bands obtained by TLC overlay with anti-Gaipi-4(Fucal-3)GlcNAcpi-R antibody indicated the presence of lipid bound Lewis" antigen, whereas the isomeric Lewis 3 structure (Gaipi-3(Fuccd-4)GlcNAcpi-R) was not detectable. G M3 substituted with Neu5Gc and Neu5Ac in a 2:1 ratio was the major ganglioside comprising about 95% within the whole ganglioside fraction. G M3 -structures were further characterized by FAB-MS and GC-MS of the native compounds and their permethylated derivatives. C 18 -sphingosine was the only long chain base, whereas variation occurred due to C^o^a and C 16 fatty acids. Terminally a2-3 sialylated neolacto-series gangliosides with nLcOse 4 -and nLcOse 6 Cer (<5% of total resorcinol stain) were found in almost equal quantities, whereas no a.2-6 sialylated counterparts were detected.

Glycosphingolipid Changes Induced by Advanced Glycation End-Products

“…Glycosphingolipids, which all migrated in doublets, were identified by their comigration with standards on the HPTLC plates. We found that the major ganglioside of BRP and BREC was GM3 (71 and 81% of the total gangliosides, respectively), which agrees with the ganglioside composition of other vascular cells such as human umbilical vein endothelial cells and smooth muscle cells (22) or bovine aortic endothelial cells (23), where GM3 is the predominant ganglioside.…”

Cell membrane GM 1 ganglioside is a functional coreceptor for fibroblast growth factor 2

“…of ganglioside biosynthesis leads to a decreased capacity of endothelial cells to respond to FGF2. Endothelial cells express neuraminic acid-bearing GM 3 and GM 1 gangliosides (24,25). GM 1 binds FGF2 with a potency significantly higher than GM 3 (13).…”

Neuronal activity-dependent membrane traffic at the neuromuscular junction

“…The TTC fragment also displays gangliosides and p15-binding properties (11)(12)(13)(14)(15). Compared with neuronal tissues, b-type gangliosides are expressed less abundantly in skeletal muscle or endothelial cells (16,17), but that could explain binding and endocytosis into these cells. Gangliosides are known to segregate into membrane lipid microdomains called rafts and͞or caveolae, which contain high amounts of glycosphingolipids and cholesterol but also are enriched in a specific signaling molecule(s) (for reviews see refs.…”