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Recombinant mucin-type proteins carrying LacdiNAc on different O-glycan core chains fail to support H. pylori binding.
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2020 (Engelska)Ingår i: Molecular Omics, E-ISSN 2515-4184, Vol. 16, nr 3, s. 243-257Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

The β4-N-acetylgalactosaminyltransferase 3 (B4GALNT3) transfers GalNAc in a β1,4-linkage to GlcNAc forming the LacdiNAc (LDN) determinant on oligosaccharides. The LacdiNAc-binding adhesin (LabA) has been suggested to mediate attachment of Helicobacter pylori to the gastric mucosa via binding to the LDN determinant. The O-glycan core chain specificity of B4GALNT3 is poorly defined. We investigated the specificity of B4GALNT3 on GlcNAc residues carried by O-glycan core 2, core 3 and extended core 1 precursors using transient transfection of CHO-K1 cells and a mucin-type immunoglobulin fusion protein as reporter protein. Binding of the LabA-positive H. pylori J99 and 26695 strains to mucin fusion proteins carrying the LDN determinant on different O-glycan core chains and human gastric mucins with and without LDN was assessed in a microtiter well-based binding assay, while the binding of 125I-LDN-BSA to various clinical H. pylori isolates was assessed in solution. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) and western blotting confirmed the requirement of a terminal GlcNAc for B4GALNT3 activity. B4GALNT3 added a β1,4-linked GalNAc to GlcNAc irrespective of whether the latter was carried by a core 2, core 3 or extended core 1 chain. No LDN-mediated adhesion of H. pylori strains 26 695 and J99 to LDN determinants on gastric mucins or a mucin-type fusion protein carrying core 2, 3 and extended core 1 O-glycans were detected in a microtiter well-based adhesion assay and no binding of a 125I-labelled LDN-BSA neoglycoconjugate to clinical H. pylori isolates was identified.

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Royal Society of Chemistry, 2020. Vol. 16, nr 3, s. 243-257
Nationell ämneskategori
Medicinska och farmaceutiska grundvetenskaper Medicinsk bioteknologi (med inriktning mot cellbiologi (inklusive stamcellsbiologi), molekylärbiologi, mikrobiologi, biokemi eller biofarmaci)
Identifikatorer
URN: urn:nbn:se:umu:diva-171214DOI: 10.1039/c9mo00175aISI: 000542588800005PubMedID: 32267274Scopus ID: 2-s2.0-85086515284OAI: oai:DiVA.org:umu-171214DiVA, id: diva2:1432989
Tillgänglig från: 2020-05-28 Skapad: 2020-05-28 Senast uppdaterad: 2023-03-24Bibliografiskt granskad

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Olofsson Edlund, JohanBoren, Thomas

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Olofsson Edlund, JohanBoren, Thomas
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Institutionen för medicinsk kemi och biofysik
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Molecular Omics
Medicinska och farmaceutiska grundvetenskaperMedicinsk bioteknologi (med inriktning mot cellbiologi (inklusive stamcellsbiologi), molekylärbiologi, mikrobiologi, biokemi eller biofarmaci)

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