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 35S-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 Galβ3GlcNAcβ3-Galβ4Glcβ1Cer (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, Galβ3GlcNAc is an essential part of the binding epitope.

Subject headings

NATURVETENSKAP  -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)

NATURAL SCIENCES  -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)

MEDICIN OCH HÄLSOVETENSKAP  -- Medicinska och farmaceutiska grundvetenskaper -- Andra medicinska och farmaceutiska grundvetenskaper (hsv//swe)

MEDICAL AND HEALTH SCIENCES  -- Basic Medicine -- Other Basic Medicine (hsv//eng)

MEDICIN OCH HÄLSOVETENSKAP  -- Medicinska och farmaceutiska grundvetenskaper -- Mikrobiologi inom det medicinska området (hsv//swe)

MEDICAL AND HEALTH SCIENCES  -- Basic Medicine -- Microbiology in the medical area (hsv//eng)

Keyword

Bacteria

Cells

Degradation

Gas chromatography

Hydrolysis

Mass spectrometry

Nuclear magnetic resonance spectroscopy

Thin-layer chromatograms

Lipids

galactose

galactosidase

glucan synthase

glycosphingolipid

lactose

lactotetraosylceramide

n acetylglucosamine

sulfur 35

tetrose

unclassified drug

article

autoradiography

bacterium adherence

controlled study

gas chromatography

Helicobacter pylori

human

human tissue

mass spectrometry

meconium

molecular model

priority journal

proton nuclear magnetic resonance

stomach epithelium

thin layer chromatography

Chromatography

Gas

Chromatography

Thin Layer

Epithelium

Epitopes

Galactose

Gastric Mucosa

Glycoside Hydrolases

Glycosphingolipids

Helicobacter pylori

Humans

Lactose

Magnetic Resonance Spectroscopy

Mass Spectrometry

Meconium

Models

Chemical

Models

Molecular

Oligosaccharides

Polysaccharides

Protein Binding

Protein Structure

Tertiary

Temperature

Thermodynamics

Bacteria (microorganisms)

Helicobacter

Helicobacter pylori

Negibacteria

Publication and Content Type

ref (subject category)

art (subject category)