Chemical and Immunological Characterization of Galactosyl-β1-3-Globoside in Bovine, Human, and Rat Brain |
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| Authors: | Somsankar Dasgupta Edward L Hogan Herman van Halbeek |
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| Institution: | Department of Neurology, Medical University of South Carolina, Charleston, South Carolina;and; Complex Carbohydrate Research Center, University of Georgia, Athens, Georgia, U.S.A. |
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| Abstract: | Abstract: 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 Galβ1-3GalNAcβ1-3Galα1-4Galβ1-4Glc\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. |
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| Keywords: | Neutral glycosphingolipids Brain Myelin NMR |
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