Chemical and immunological characterization of galactosyl-beta 1-3-globoside in bovine, human, and rat brain.
Our studies of bovine brain neutral glycosphingolipids (Ngsls) have revealed the presence of several short-chain (containing -CHO 1-4) and previously uncharacterized long-chain (-CHO > 4-5) Ngsls. We reported the structural characterization of brain GgOse4Cer (GA1) and have now purified another brain Ngsl to homogeneity. The purified Ngsl migrated close to standard GgOse4Cer and nLcOse5Cer on a TLC plate employing two different solvent systems. The carbohydrate molar composition indicated the presence of Gal/Glc/GalNAc in a ratio of 2.8:1.0:0.9. Five permethylated alditol acetate peaks were characterized as 2,3,4,6-OMe4Gal, 2,4,6-OMe3Gal, 2,3,6-OMe3Gal, 2,3,6-OMe3Glc, and 4,6-OMe2GalNAcMe by gas chromatography-mass spectrometry. The anomeric carbohydrate sequence has been determined by specific exoglycosidase digestion. Six-hundred megahertz 1H NMR spectroscopy of the oligosaccharide released by ceramide glycanase hydrolysis confirmed the structure of the Ngsl as Gal beta 1-3GalNAc beta 1-3Gal alpha 1-4Gal beta 1-4Glc beta 1-1Cer or IV3GalGbOse4Cer. Using the immunooverlay technique with anti-stage-specific embryonic antigen 3 antibody, it was found in bovine, rat, and normal adult human brain and bovine myelin, but not in human or rat myelin.
Dasgupta
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S
Department of Neurology, Medical University of South Carolina, Charleston 29425-2232, USA.
Hogan
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EL
van Halbeek
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H
D000328:Adult; D000818:Animals; D000911:Antibodies, Monoclonal; D015295:Antigens, Tumor-Associated, Carbohydrate; D001921:Brain / Q000378:metabolism*; D002240:Carbohydrate Sequence; D002417:Cattle; D002855:Chromatography, Thin Layer; D008401:Gas Chromatography-Mass Spectrometry; D006028:Glycosphingolipids / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D007158:Immunologic Techniques; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009186:Myelin Sheath / Q000378:metabolism; D051381:Rats; D055746:Stage-Specific Embryonic Antigens
D000911:Antibodies, Monoclonal; D015295:Antigens, Tumor-Associated, Carbohydrate; D006028:Glycosphingolipids; D055746:Stage-Specific Embryonic Antigens; C052924:stage-specific embryonic antigen-3