Characterization of Gangliosides of Epithelial Cells of Calf Small Intestine, with Special Reference to Receptor-Active Sequences for Enteropathogenic Escherichia coli K99

• Published in: Journal of biochemistry (Tokyo) 1994-09, Vol.116 (3), p.560-571
• Main Authors: Teneberg, Susann, Willemsen, Peter T.J., de Graaf, Frits K., Stenhagen, Gunnar, Pimlott, Weston, Jovall, Per-Åke, Ångström, Jonas, Karlsson, Karl-Anders
• Format: Article
• Language: English
• Subjects: Animals; bacterial adhesion; calf small intestine; Carbohydrate Sequence; Cattle; Chromatography, Thin Layer; E. coli K99; Epithelial Cells; Epithelium - metabolism; Epitopes; Escherichia coli; Escherichia coli - metabolism; ganglioside receptors; Gangliosides - metabolism; Intestinal Mucosa - cytology; Intestinal Mucosa - metabolism; Intestine, Small - cytology; Intestine, Small - metabolism; Magnetic Resonance Spectroscopy - methods; Mass Spectrometry; Molecular Sequence Data; N-glycolylneuraminic acid; Protons; Receptors, Cell Surface - metabolism; Reference Values
• ISSN: 0021-924X
• DOI: 10.1093/oxfordjournals.jbchem.a124562

Glycolipids were prepared from epithelial cells of the small intestine of a newborn calf and assayed for Escherichia coli K99 binding activity on thin-layer chromatograms and in microtiter wells. The bacteria did not bind to any of the non-acid glycolipids, while in the acid fraction several binding-positive glycolipids were detected. The acid glycolipids were isolated and characterized by mass spectrometry, proton NMR spectroscopy and other methods. The following gangliosides were identified, mainly from the epithelial cells from the upper part of the small intestine: NeuAcα2–3Galβl-4Glcβl-Cer (NeuAc-GM3), NeuGcα2–3Galβl-4Glcβl-Cer (NeuGc-GM3), GalNAcβl-4(NeuGcα2–3)Galβl-4Glcβl-Cer (NeuGc-GM2), Galβl-3GalNAcβl-4(NeuGcα2–3)Galβl-4Glcαl-Cer (NeuGc-GMl), and NeuGcα2–3Galβl-3GalNAcβl-4(NeuGcα2–3)Galβl-4Glcβl-Cer (NeuGc-GDla). A positive binding was demonstrated to NeuGc-GM3, NeuGc-GM2, and NeuGc-GDla, while NeuAc-GM3 and NeuGc-GMl were negative. The binding pattern differed somewhat for total acid glycolipids of epithelial cells from three different parts of the small intestine. Based on binding preferences of E. coli K99 to a number of glycolipids of various origins, in comparison with calculated minimum energy conformations, a binding epitope was delineated.