A novel glycosphingolipid expressed in pig kidney: Gal alpha 1-3Lewis(x) hexaglycosylceramide.
Immunodetection of thin layer chromatograms of neutral glycosphingolipids of pig kidney cortex with a polyclonal antibody directed against the Gal alpha 1-3Gal 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 Gal alpha 1-3Gal beta 1-4(Fuc alpha 1-3)GlcNAc beta 1-3Gal beta 1-4Glc beta 1-Cer by fast atom bombardment- and desorption-chemical ionization-mass spectrometry, methylation analysis and hydrolysis with alpha-galactosidase followed by immunostaining with an anti-Lewis(x) 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 Gal alpha 1-3 Lewis(x) determinant appeared to be developmentally regulated as it increased with age. The characterization of Gal alpha 1-3Le(x) 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 Le(x)-based glycolipids.
Bouhours
D
D
Institut de Transplantation et de Recherche en Transplantation, INSERM U.437, Centre Hospitalier Universitaire, Nantes, France.
Liaigre
J
J
Naulet
J
J
Maume
D
D
Bouhours
J F
JF
D000818:Animals; D002240:Carbohydrate Sequence; D002518:Ceramides; D002645:Chickens; D002855:Chromatography, Thin Layer / Q000379:methods; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000737:chemistry; D006028:Glycosphingolipids / Q000737:chemistry* / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D007668:Kidney / Q000737:chemistry* / Q000378:metabolism; D007983:Lewis Blood Group Antigens / Q000737:chemistry* / Q000276:immunology*; D016256:Lewis X Antigen / Q000031:analogs & derivatives; D009682:Magnetic Resonance Spectroscopy; D008745:Methylation; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry* / Q000276:immunology* / Q000378:metabolism; D016339:Spectrometry, Mass, Fast Atom Bombardment
D002518:Ceramides; D000939:Epitopes; C106297:Gal-1-3-Lewis(x) hexaglycosylceramide; D006028:Glycosphingolipids; D007983:Lewis Blood Group Antigens; D016256:Lewis X Antigen; D009844:Oligosaccharides