Lactotetraosylceramide, a novel glycosphingolipid receptor for Helicobacter pylori, present in human gastric epithelium.
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.
Teneberg
Susann
S
Institute of Medical Biochemistry, Göteborg University, P. O. Box 440, SE 405 30 Göteborg, Sweden. Susann.Teneberg@medkem.gu.se
Leonardsson
Irene
I
Karlsson
Hasse
H
Jovall
Per-Ake
PA
Angstrom
Jonas
J
Danielsson
Dan
D
Naslund
Ingmar
I
Ljungh
Asa
A
Wadstrom
Torkel
T
Karlsson
Karl-Anders
KA
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
D000939:Epitopes; D006028:Glycosphingolipids; D009844:Oligosaccharides; D011134:Polysaccharides; C458510:lactotetraosylceramide; D006026:Glycoside Hydrolases; C051111:ceramide glycanase; C020668:glycanase; D007785:Lactose; D005690:Galactose