title: Lactotetraosylceramide, a novel glycosphingolipid receptor for Helicobacter pylori, present in human gastric epithelium.
abstract: The binding of Helicobacter pylori to glycosphingolipids was examined by binding of (35)S-labeled bacteria to glycosphingolipids on thin-layer chromatograms. In addition to previously reported binding specificities, a selective binding to a non-acid tetraglycosylceramide of human meconium was found. This H. pylori binding glycosphingolipid was isolated and, on the basis of mass spectrometry, proton NMR spectroscopy, and degradation studies, were identified as Galbeta3GlcNAcbeta3Galbeta4Glcbeta1Cer (lactotetraosylceramide). When using non-acid glycosphingolipid preparations from human gastric epithelial cells, an identical binding of H. pylori to the tetraglycosylceramide interval was obtained in one of seven samples. Evidence for the presence of lactotetraosylceramide in the binding-active interval was obtained by proton NMR spectroscopy of intact glycosphingolipids and by gas chromatography-electron ionization mass spectrometry of permethylated tetrasaccharides obtained by ceramide glycanase hydrolysis. The lactotetraosylceramide binding property was detected in 65 of 74 H. pylori isolates (88%). Binding of H. pylori to lactotetraosylceramide on thin-layer chromatograms was inhibited by preincubation with lactotetraose but not with lactose. Removal of the terminal galactose of lactotetraosylceramide by galactosidase hydrolysis abolished the binding as did hydrazinolysis of the acetamido group of the N-acetylglucosamine. Therefore, Galbeta3GlcNAc is an essential part of the binding epitope.
authors.affiliation: Institute of Medical Biochemistry, Göteborg University, P. O. Box 440, SE 405 30 Göteborg, Sweden. Susann.Teneberg@medkem.gu.se
authors.forename: Susann
authors.initials: S
authors.lastname: Teneberg
authors.forename: Irene
authors.initials: I
authors.lastname: Leonardsson
authors.forename: Hasse
authors.initials: H
authors.lastname: Karlsson
authors.forename: Per-Ake
authors.initials: PA
authors.lastname: Jovall
authors.forename: Jonas
authors.initials: J
authors.lastname: Angstrom
authors.forename: Dan
authors.initials: D
authors.lastname: Danielsson
authors.forename: Ingmar
authors.initials: I
authors.lastname: Naslund
authors.forename: Asa
authors.initials: A
authors.lastname: Ljungh
authors.forename: Torkel
authors.initials: T
authors.lastname: Wadstrom
authors.forename: Karl-Anders
authors.initials: KA
authors.lastname: Karlsson
mesh_terms: D002849:Chromatography, Gas; D002855:Chromatography, Thin Layer; D004848:Epithelium / Q000378:metabolism*; D000939:Epitopes; D005690:Galactose / Q000378:metabolism; D005753:Gastric Mucosa / Q000378:metabolism*; D006026:Glycoside Hydrolases / Q000378:metabolism; D006028:Glycosphingolipids / Q000737:chemistry* / Q000378:metabolism*; D016480:Helicobacter pylori / Q000378:metabolism*; D006801:Humans; D007785:Lactose / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D008470:Meconium / Q000378:metabolism; D008956:Models, Chemical; D008958:Models, Molecular; D009844:Oligosaccharides; D011134:Polysaccharides / Q000737:chemistry; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D013696:Temperature; D013816:Thermodynamics
chemical_list: D000939:Epitopes; D006028:Glycosphingolipids; D009844:Oligosaccharides; D011134:Polysaccharides; C458510:lactotetraosylceramide; D006026:Glycoside Hydrolases; C051111:ceramide glycanase; C020668:glycanase; D007785:Lactose; D005690:Galactose