Open AccessNeu5Acα3Gal is part of the Helicobacter pylori binding epitope in polyglycosylceramides of human erythrocytes
Author(s) -
Johansson Lena,
Johansson Petra,
MillerPodraza Halina
Publication year - 1999
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1046/j.1432-1327.1999.00893.x
Subject(s) - sialic acid , chemistry , molecular mass , biochemistry , helicobacter pylori , chromatography , biology , enzyme , genetics
The sialic acid dependent binding by the human pathogen Helicobacter pylori to polyglycosylceramides of human erythrocytes was investigated. Polyglycosylceramides, complex glycosphingolipids with a branched N ‐acetyllactosamine core, were isolated from human erythrocytes, blood group O, and subfractionated after peracetylation by anion‐exchange chromatography. Three subfractions were deacetylated, analysed by matrix‐assisted laser desorption ionization‐time of flight MS and 2D 1 H NMR spectroscopy. The observed mass ranges were m / z = 3093–7622, 3968–7255 and 3459–7987 in the mass spectra of the first, second and third fractions, respectively. The observed ions agreed with the general formula Hex ( x +2) HexNAc x Fuc y Neu5Ac z Cer. Two‐dimensional 1 H total correlation spectra of the mixtures showed that the first fraction contained 3‐linked sialic acid and the second and third fractions contained both 3‐linked and 6‐linked sialic acid. Thin‐layer chromatogram binding assays using the lectins from Maackia amurensis , specific for Neu5Acα3Galβ4GlcNAc, and Sambucus nigra , specific for Neu5Acα6Gal/GalNAc, were used to confirm this distribution. H. pylori recognized all three fractions in the binding assay, indicating that the 3‐linked, rather than 6‐linked, sialic acid is essential for binding.