Molecular characterization and immunohistochemical localization of IV(4)GalNAcGgOse(4)Cer: a naturally occurring novel neutral glycosphingolipid in bovine brain.

摘要

A pair of novel neutral glycosphingolipids (Ngsls) has been identified in bovine brain. Their mobilities on thin layer chromatography were slightly different from a standard pentaglycosylceramide (nLcOse(5)Cer from bovine erythrocytes). The compounds were purified to homogeneity by column chromatography. Their fatty acid and base compositions, their monosaccharide compositions and sugar linkage positions were determined by gas-liquid chromato-graphy/mass spectrometry. Carbohydrate sequence analy-sis by(1)H NMR spectroscopy and stepwise exoglyco-sidase digestion indicated the following pentaglycosyl structure for the oligosaccharide moiety of both Ngsls: GalNAcbeta1-4Galbeta1-3GalNAcbeta1-4Galbeta1-4Gl c. The two Ngsls (abbreviated as IV(4)GalNAcGgOse(4)Cer or GalNAc-GA1), differ in their ceramide compositions, having d18:0 and d18:1 sphingosine as their long chain bases. A monospecific polyclonal anti-GalNAc-GA1 antibody, prepared in rabbit and purified by affinity chromatography, stained the neurons of cerebral cortex and cerebellum including Purkinje cells in adult rat brain, indicating that the novel GalNAc-GA1 is associated with cerebellar and other neurons in vertebrate central nervous system.