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Epitope mapping of histo blood group antigens bound to norovirus VLPs using STD NMR experiments reveals fine details of molecular recognition

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Abstract

Attachment of human noroviruses to histo blood group antigens (HBGAs) is thought to be critical for the infection process. Therefore, we have determined binding epitopes of synthetic type 1 to 6 blood group A- and B-tetrasaccharides binding to GII.4 human Norovirus virus like particles (VLPs) using STD NMR experiments. So far, little information is available from crystal structure analysis studies on the interactions of the reducing-end sugars with the protruding domain (P-domain) of the viral coat protein VP1. Here, we show that the reducing-end sugars make notable contacts with the protein surface. The type of glycosidic linkage, and the identity of the sugar at the reducing end modulate HBGA recognition. Most strikingly, type 2 structures yield only very poor saturation transfer indicating impeded binding. This observation is in accordance with previous mass spectrometry based affinity measurements, and can be understood based on recent crystal structure data of a complex of highly homologous GII.4 P-dimers with H-type 2 trisaccharide where the N-acetyl group of the reducing N-acetyl glucosamine residue points towards a loop comprising amino acids Q390 to H395. We suggest that in our case, binding of type 2 A- and B-tetrasaccharides leads to steric conflicts with this loop. In order to identify factors determining L-Fuc recognition, we also synthesized GII.4 VLPs with point mutations D391A and H395A. Prior studies had suggested that these residues, located in a second shell around the L-Fuc binding site, assist L-Fuc binding. STD NMR experiments with L-Fuc and B-trisaccharide in the presence of wild type and mutant VLPs yield virtually identical binding epitopes suggesting that these two mutations do not significantly alter HBGA recognition. Our study emphasizes that recognition of α−(1→2)-linked L-Fuc residues is a conserved feature of GII.4 noroviruses. However, structural variation of the HBGA core structures clearly modulates molecular recognition depending on the genotype.

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References

  1. Ahmed, S.M., Hall, A.J., Robinson, A.E., Verhoef, L., Premkumar, P., Parashar, U.D., Koopmans, M., Lopman, B.A.: Global prevalence of norovirus in cases of gastroenteritis: a systematic review and meta-analysis. Lancet Infect. Dis. 14(8), 725–730 (2014). doi:10.1016/s1473-3099(14)70767-4

    Article  PubMed  Google Scholar 

  2. Lee, B.Y., McGlone, S.M., Bailey, R.R., Wettstein, Z.S., Umscheid, C.A., Muder, R.R.: Economic impact of outbreaks of norovirus infection in hospitals. Infect. Control Hosp. Epidemiol: the official journal of the Society of Hospital Epidemiologists of America. 32(2), 191–193 (2011). doi:10.1086/657910

    Article  Google Scholar 

  3. Koo, H.L., Ajami, N., Atmar, R.L., DuPont, H.L.: Noroviruses: the principal cause of foodborne disease worldwide. Discov. Med. 10(50), 61–70 (2010)

    PubMed  PubMed Central  Google Scholar 

  4. Glass, R.I., Parashar, U.D., Estes, M.K.: Norovirus gastroenteritis. N. Engl. J. Med. 361, 1776–1785 (2009)

    Article  CAS  PubMed  Google Scholar 

  5. Tan, M., Jiang, X.: Norovirus-host interaction: implications for disease control and prevention. Expert Rev. Mol. Med. 9(19), 1–22 (2007). doi:10.1017/S1462399407000348

    Article  PubMed  Google Scholar 

  6. Lindesmith, L., Moe, C., Marionneau, S., Ruvoen, N., Jiang, X., Lindblad, L., Stewart, P., LePendu, J., Baric, R.: Human susceptibility and resistance to Norwalk virus infection. Nat. Med. 9(5), 548–553 (2003). doi:10.1038/nm860

    Article  CAS  PubMed  Google Scholar 

  7. Huang, P., Farkas, T., Marionneau, S., Zhong, W., Ruvoen-Clouet, N., Morrow, A.L., Altaye, M., Pickering, L.K., Newburg, D.S., LePendu, J., Jiang, X.: Noroviruses bind to human ABO, Lewis, and secretor histo-blood group antigens: identification of 4 distinct strain-specific patterns. J Infect Dis. 188(1), 19–31 (2003). doi:10.1086/375742

    Article  CAS  PubMed  Google Scholar 

  8. Nystrom, K., Le Gall-Recule, G., Grassi, P., Abrantes, J., Ruvoen-Clouet, N., Le Moullac-Vaidye, B., Lopes, A.M., Esteves, P.J., Strive, T., Marchandeau, S., Dell, A., Haslam, S.M., Le Pendu, J.: Histo-blood group antigens act as attachment factors of rabbit hemorrhagic disease virus infection in a virus strain-dependent manner. PLoS Pathog. 7(8), e1002188 (2011). doi:10.1371/journal.ppat.1002188

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Bartnicki, E., Cunha, J.B., Kolawole, A.O., Wobus, C.E.: Recent advances in understanding noroviruses. F1000Research. 6, 1–9 (2017). doi:10.12688/f1000research.10081.1

    Article  Google Scholar 

  10. Schroten, H., Hanisch, F.G., Hansman, G.S.: Human norovirus interactions with Histo-blood group antigens and human milk oligosaccharides. J. Virol. 90(13), 5855–5859 (2016). doi:10.1128/JVI.00317-16

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Prasad, B.V., Hardy, M.E., Dokland, T., Bella, J., Rossmann, M.G., Estes, M.K.: X-ray crystallographic structure of the Norwalk virus capsid. Science. 286(5438), 287–290 (1999)

    Article  CAS  PubMed  Google Scholar 

  12. Chen, R., Neill, J.D., Estes, M.K., Prasad, B.V.: X-ray structure of a native calicivirus: structural insights into antigenic diversity and host specificity. Proc. Natl. Acad. Sci. U. S. A. 103(21), 8048–8053 (2006). doi:10.1073/pnas.0600421103

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Cao, S., Lou, Z., Tan, M., Chen, Y., Liu, Y., Zhang, Z., Zhang, X.C., Jiang, X., Li, X., Rao, Z.: Structural basis for the recognition of blood group trisaccharides by norovirus. J. Virol. 81(11), 5949–5957 (2007). doi:10.1128/JVI.00219-07

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. de Graaf, M., van Beek, J., Koopmans, M.P.: Human norovirus transmission and evolution in a changing world. Nat. Rev. Microbiol. 14(7), 421–433 (2016). doi:10.1038/nrmicro.2016.48

    Article  PubMed  Google Scholar 

  15. Tan, M., Xia, M., Chen, Y., Bu, W., Hegde, R.S., Meller, J., Li, X., Jiang, X.: Conservation of carbohydrate binding interfaces: evidence of human HBGA selection in norovirus evolution. PLoS ONE. 4(4), e5058 (2009). doi:10.1371/journal.pone.0005058

    Article  PubMed  PubMed Central  Google Scholar 

  16. Singh, B.K., Leuthold, M.M., Hansman, G.S.: Human noroviruses' fondness for histo-blood group antigens. J. Virol. 89(4), 2024–2040 (2015). doi:10.1128/JVI.02968-14

    Article  PubMed  Google Scholar 

  17. Hansman, G.S., Taylor, D.W., McLellan, J.S., Smith, T.J., Georgiev, I., Tame, J.R., Park, S.Y., Yamazaki, M., Gondaira, F., Miki, M., Katayama, K., Murata, K., Kwong, P.D.: Structural basis for broad detection of genogroup II noroviruses by a monoclonal antibody that binds to a site occluded in the viral particle. J. Virol. 86(7), 3635–3646 (2012). doi:10.1128/JVI.06868-11

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Shanker, S., Choi, J.M., Sankaran, B., Atmar, R.L., Estes, M.K., Prasad, B.V.: Structural analysis of histo-blood group antigen binding specificity in a norovirus GII.4 epidemic variant: implications for epochal evolution. J. Virol. 85(17), 8635–8645 (2011). doi:10.1128/JVI.00848-11

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Fiege, B., Rademacher, C., Cartmell, J., Kitov, P.I., Parra, F., Peters, T.: Molecular details of the recognition of blood group antigens by a human norovirus as determined by STD NMR spectroscopy. Angew. Chem. 51(4), 928–932 (2012). doi:10.1002/anie.201105719

    Article  CAS  Google Scholar 

  20. Mallagaray, A., Rademacher, C., Parra, F., Hansman, G., Peters, T.: STD NMR titrations reveal complex multistep-binding of L-Fucose to norovirus particles. Glycobiology. 27(1), 80–86 (2017). doi:10.1093/glycob/cww070

    Article  PubMed  Google Scholar 

  21. Mallagaray, A., Lockhauserbäumer, J., Hansman, G.S., Uetrecht, C., Peters, T.: Attachment of norovirus to Histo blood group antigens: a cooperative multistep process. Angew. Chem. Int. Ed. 54(41), 12014–12019 (2015)

    Article  CAS  Google Scholar 

  22. Koromyslova, A.D., Leuthold, M.M., Bowler, M.W., Hansman, G.S.: The sweet quartet: binding of fucose to the norovirus capsid. Virology. 483, 203–208 (2015). doi:10.1016/j.virol.2015.04.006

    Article  CAS  PubMed  Google Scholar 

  23. Han, L., Kitova, E.N., Tan, M., Jiang, X., Pluvinage, B., Boraston, A.B., Klassen, J.S.: Affinities of human histo-blood group antigens for norovirus capsid protein complexes. Glycobiology. 25, 170–180 (2015). doi:10.1093/glycob/cwu100

    Article  CAS  PubMed  Google Scholar 

  24. Mayer, M., Meyer, B.: Group epitope mapping by saturation transfer difference NMR to identify segments of a ligand in direct contact with a protein receptor. J. Am. Chem. Soc. 123, 6108–6117 (2001)

    Article  CAS  PubMed  Google Scholar 

  25. Mayer, M., Meyer, B.: Characterization of ligand binding by saturation transfer difference NMR spectroscopy. Angew. Chem. Int. Ed. 38(12), 1784–1787 (1999)

    Article  CAS  Google Scholar 

  26. Meloncelli, P.J., West, L.J., Lowary, T.L.: Synthesis and NMR studies on the ABO histo-blood group antigens: synthesis of type III and IV structures and NMR characterization of type I-VI antigens. Carbohydr. Res. 346(12), 1406–1426 (2011). doi:10.1016/j.carres.2011.03.008

    Article  CAS  PubMed  Google Scholar 

  27. Meloncelli, P.J., Lowary, T.L.: Synthesis of ABO histo-blood group type I and II antigens. Carbohydr. Res. 345(16), 2305–2322 (2010). doi:10.1016/j.carres.2010.08.012

    Article  CAS  PubMed  Google Scholar 

  28. Meloncelli, P.J., Lowary, T.L.: Synthesis of ABO Histo-blood group type V and VI antigens. Aust. J. Chem. 62, 558–574 (2009)

    Article  CAS  Google Scholar 

  29. Liu, W., Chen, Y., Jiang, X., Xia, M., Yang, Y., Tan, M., Li, X., Rao, Z.: A unique human norovirus lineage with a distinct HBGA binding Interface. PLoS Pathog. 11(7), e1005025 (2015). doi:10.1371/journal.ppat.1005025

    Article  PubMed  PubMed Central  Google Scholar 

  30. Singh, B.K., Leuthold, M.M., Hansman, G.S.: Structural constraints on human norovirus binding to histo-blood group antigens. mSphere. 1(2), (2016). doi:10.1128/mSphere.00049-16

  31. Pengelley, S.C., Chapman, D.C., Mark Abbott, W., Lin, H.H., Huang, W., Dalton, K., Jones, I.M.: A suite of parallel vectors for baculovirus expression. Protein Expr. Purif. 48(2), 173–181 (2006). doi:10.1016/j.pep.2006.04.016

    Article  CAS  PubMed  Google Scholar 

  32. Zhao, Y., Chapman, D.A.G., Jones, I.M.: Improving baculovirus recombination. Nucleic Acids Res. 31(2), 6e-6 (2003). doi:10.1093/nar/gng006

    Article  Google Scholar 

  33. Kirschner, K.N., Yongye, A.B., Tschampel, S.M., Gonzalez-Outeirino, J., Daniels, C.R., Foley, B.L., Woods, R.J.: GLYCAM06: a generalizable biomolecular force field. Carbohydrates. J. Comput. Chem. 29(4), 622–655 (2008). doi:10.1002/jcc.20820

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Tan, M., Xia, M., Cao, S., Huang, P., Farkas, T., Meller, J., Hegde, R.S., Li, X., Rao, Z., Jiang, X.: Elucidation of strain-specific interaction of a GII-4 norovirus with HBGA receptors by site-directed mutagenesis study. Virology. 379(2), 324–334 (2008). doi:10.1016/j.virol.2008.06.041

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Nasir, W., Frank, M., Kunze, A., Bally, M., Parra, F., Nyholm, P.G., Hook, F., Larson, G.: Histo-blood group antigen presentation is critical for norovirus VLP binding to glycosphingolipids in model membranes. ACS Chem. Biol. (2017). doi:10.1021/acschembio.7b00152

  36. Rademacher, C., Krishna, N.R., Palcic, M., Parra, F., Peters, T.: NMR experiments reveal the molecular basis of receptor recognition by a calicivirus. J. Am. Chem. Soc. 130(11), 3669–3675 (2008)

    Article  CAS  PubMed  Google Scholar 

  37. Jiang, X., Wang, M., Graham, D.Y., Estes, M.K.: Expression, self-assembly, and antigenicity of the Norwalk virus capsid protein. J. Virol. 66(11), 6527–6532 (1992)

    CAS  PubMed  PubMed Central  Google Scholar 

  38. Rademacher, C., Guiard, J., Kitov, P.I., Fiege, B., Dalton, K.P., Parra, F., Bundle, D.R., Peters, T.: Targeting norovirus infection-multivalent entry inhibitor design based on NMR experiments. Chemistry. 17(27), 7442–7453 (2011). doi:10.1002/chem.201003432

    Article  CAS  PubMed  Google Scholar 

  39. Rademacher, C., Peters, T.: Molecular recognition of ligands by native viruses and virus-like particles as studied by NMR experiments. In: Peters, T. (ed.) Topics in Current Chemistry, Bioactive Conformation II, Vol. 273. Bioactive Conformation II, pp. 183–202. Springer, Berlin Heidelberg (2008)

    Google Scholar 

  40. Mayer, M., James, T.L.: NMR-based characterization of phenothiazines as a RNA binding scaffold. J. Am. Chem. Soc. 126(13), 4453–4460 (2004). doi:10.1021/ja0398870

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

Financial support from the DFG (PE494/8-1 and PE494,12-1 with the research unit FOR2327) is gratefully acknowledged. The authors thank the University of Lübeck and the Alberta Glycomics Centre for financial support. The DFG and the state of Schleswig-Holstein are thanked for a grant for the cryogenic probe (HBFG 101/192-1). FP and KPD were supported by grant GRUPIN14-099 from Principado de Asturias (Spain), cofinanced by FEDER. Prof. Monica Palcic (University of Victoria, BC, Canada) is thanked for a kind gift of the blood group B-trisaccharide. Dr. Alvaro Mallagaray from the University of Lübeck and Dr. Grant Hansman from the DKFZ in Heidelberg are thanked for stimulating discussions. Dr. Thorsten Biet, Thies Köhli, and Wilfried Hellebrandt are thanked for technical assistance.

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Author notes

  1. Brigitte Fiege

    Present address: Institute of Molecular Pharmacy, Pharmacenter, University of Basel, Klingelbergstrasse 50, 4056, Basel, Switzerland

  2. Mila Leuthold

    Present address: Molecular Virology, Heidelberg University Hospital, Im Neuenheimer Feld 345, 69120, Heidelberg, Germany

  3. Brigitte Fiege and Mila Leuthold contributed equally to this study.

Authors and Affiliations

  1. Center of Structural and Cell Biology in Medicine, Institute of Chemistry and Metabolomics, University of Luebeck, Ratzeburger Allee 160, 23562, Luebeck, Germany

    Brigitte Fiege, Mila Leuthold & Thomas Peters

  2. Instituto Universitario de Biotecnología de Asturias, Departamento de Bioquímica y Biología Molecular, Universidad de Oviedo, 33006, Oviedo, Spain

    Francisco Parra & Kevin P. Dalton

  3. Alberta Glycomics Centre and Department of Chemistry, Gunning-Lemieux Chemistry Centre, The University of Alberta, 11227 Saskatchewan Drive, Edmonton, AB, T6G G2, Canada

    Peter J. Meloncelli & Todd L. Lowary

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  1. Brigitte Fiege
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Correspondence to Thomas Peters.

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Fiege, B., Leuthold, M., Parra, F. et al. Epitope mapping of histo blood group antigens bound to norovirus VLPs using STD NMR experiments reveals fine details of molecular recognition. Glycoconj J 34, 679–689 (2017). https://doi.org/10.1007/s10719-017-9792-5

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