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A novel glycosphingolipid expressed in pig kidney: Galα1-3Lewisx hexaglycosylceramide

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Abstract

Immunodetection of thin layer chromatograms of neutral glycosphingolipids of pig kidney cortex with a polyclonal antibody directed against the Galα1-3Gal determinant revealed several glycosphingolipids reacting with different intensities. A minor glycosphingolipid was isolated by preparative high performance thin layer chromatography. It was characterized as a type 2 hexaglycosylceramide with the following structure Galα1-3Galβ1-4(Fucα1-3)GlcNAcβ1-3Galβ1-4Glcβ1-Cer by fast atom bombardment- and desorption-chemical ionization-mass spectrometry, methylation analysis and hydrolysis with α-galactosidase followed by immunostaining with an anti-Lewisx monoclonal antibody. The proton NMR spectrum was found compatible with the proposed structure. Two other glycosphingolipids carrying the new determinant were partially characterized as an octa- and a branched-dodecaglycosylceramide. The expression of the Galα1-3Lewisx determinant appeared to be developmentally regulated as it increased with age. The characterization of Galα1-3Lex in pig kidney indicates a new epitope capable of recognition by human natural antibodies in the context of xenotransplantation of pig organs to man. It also adds new members to the family of Lex-based glycolipids. Abbreviations: HPTLC, high performance thin layer chromatography; FAB-MS, fast atom bombardment mass spectrometry; DCI, desorption-chemical ionization; Me2SO-d6, hexadeuterated dimethyl sulfoxide

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Authors and Affiliations

  1. Institut de Transplantation et de Recherche en Transplantation, INSERM U.437, Centre Hospitalier Universitaire, F-44035, Nantes Cedex, France

    Daniele Bouhours, Jerome Liaigre, Jeanne Naulet & Jean-Francois Bouhours

  2. Laboratoire de Biochimie, Ecole Nationale Veterinaire Nantes, France

    Daniel Maume

Authors
  1. Daniele Bouhours
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  2. Jerome Liaigre
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  3. Jeanne Naulet
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  4. Daniel Maume
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  5. Jean-Francois Bouhours
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Bouhours, D., Liaigre, J., Naulet, J. et al. A novel glycosphingolipid expressed in pig kidney: Galα1-3Lewisx hexaglycosylceramide. Glycoconj J 14, 29–38 (1997). https://doi.org/10.1023/A:1018504813642

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