title: Histo-Blood Group Antigen Presentation Is Critical for Binding of Norovirus VLP to Glycosphingolipids in Model Membranes.
abstract: Virus entry depends on biomolecular recognition at the surface of cell membranes. In the case of glycolipid receptors, these events are expected to be influenced by how the glycan epitope close to the membrane is presented to the virus. This presentation of membrane-associated glycans is more restricted than that of glycans in solution, particularly because of orientational constraints imposed on the glycolipid through its lateral interactions with other membrane lipids and proteins. We have developed and employed a total internal reflection fluorescence microscopy-based binding assay and a scheme for molecular dynamics (MD) membrane simulations to investigate the consequences of various glycan presentation effects. The system studied was histo-blood group antigen (HBGA) epitopes of membrane-bound glycosphingolipids (GSLs) derived from small intestinal epithelium of humans (type 1 chain) and dogs (type 2 chain) interacting with GII.4 norovirus-like particles. Our experimental results showed strong binding to all lipid-linked type 1 chain HBGAs but no or only weak binding to the corresponding type 2 chain HBGAs. This is in contrast to results derived from STD experiments with free HBGAs in solution where binding was observed for Lewis x. The MD data suggest that the strong binding to type 1 chain glycolipids was due to the well-exposed (1,2)-linked α-l-Fucp and (1,4)-linked α-l-Fucp residues, while the weaker binding or lack of binding to type 2 chain HBGAs was due to the very restricted accessibility of the (1,3)-linked α-l-Fucp residue when the glycolipid is embedded in a phospholipid membrane. Our results not only contribute to a general understanding of protein-carbohydrate interactions on model membrane surfaces, particularly in the context of virus binding, but also suggest a possible role of human intestinal GSLs as potential receptors for norovirus uptake.
authors.affiliation: Department of Clinical Chemistry and Transfusion Medicine, Sahlgrenska Academy, University of Gothenburg , Gothenburg, Sweden.
authors.forename: Waqas
authors.initials: W
authors.lastname: Nasir
authors.affiliation: Biognos AB , Generatorsgatan 1, P.O. Box 8963, 40274 Gothenburg, Sweden.
authors.forename: Martin
authors.initials: M
authors.lastname: Frank
authors.affiliation: Department of Applied Physics, Chalmers University of Technology , S-412 96 Gothenburg, Sweden.
authors.forename: Angelika
authors.identifier: 0000-0002-7887-2736
authors.initials: A
authors.lastname: Kunze
authors.affiliation: Department of Applied Physics, Chalmers University of Technology , S-412 96 Gothenburg, Sweden.
authors.forename: Marta
authors.initials: M
authors.lastname: Bally
authors.affiliation: Instituto Universitario de Biotecnología de Asturias, Departamento de Bioquimíca y Biología Molecular, Universidad de Oviedo , 33006 Oviedo, Spain.
authors.forename: Francisco
authors.initials: F
authors.lastname: Parra
authors.affiliation: Biognos AB , Generatorsgatan 1, P.O. Box 8963, 40274 Gothenburg, Sweden.|Department of Medical Biochemistry and Cell Biology, University of Gothenburg , Gothenburg, Sweden.
authors.forename: Per-Georg
authors.initials: PG
authors.lastname: Nyholm
authors.affiliation: Department of Applied Physics, Chalmers University of Technology , S-412 96 Gothenburg, Sweden.
authors.forename: Fredrik
authors.identifier: 0000-0003-1994-5015
authors.initials: F
authors.lastname: Höök
authors.affiliation: Department of Clinical Chemistry and Transfusion Medicine, Sahlgrenska Academy, University of Gothenburg , Gothenburg, Sweden.
authors.forename: Göran
authors.identifier: 0000-0002-2616-0366
authors.initials: G
authors.lastname: Larson
mesh_terms: D000818:Animals; D017951:Antigen Presentation / Q000502:physiology*; D001789:Blood Group Antigens / Q000276:immunology*; D004285:Dogs; D006028:Glycosphingolipids / Q000378:metabolism*; D006801:Humans; D007413:Intestinal Mucosa; D056004:Molecular Dynamics Simulation; D029322:Norovirus / Q000502:physiology*; D053585:Virus Attachment; D053586:Virus Internalization
chemical_list: D001789:Blood Group Antigens; D006028:Glycosphingolipids