Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

The GD1a glycan is a cellular receptor for adenoviruses causing epidemic keratoconjunctivitis

Abstract

Adenovirus type 37 (Ad37) is a leading cause of epidemic keratoconjunctivitis (EKC)1,2, a severe and highly contagious ocular disease. Whereas most other adenoviruses infect cells by engaging CD46 or the coxsackie and adenovirus receptor (CAR), Ad37 binds previously unknown sialic acid–containing cell surface molecules3,4. By glycan array screening, we show here that the receptor-recognizing knob domain of the Ad37 fiber protein specifically binds a branched hexasaccharide that is present in the GD1a ganglioside and that features two terminal sialic acids. Soluble GD1a glycan and GD1a-binding antibodies efficiently prevented Ad37 virions from binding and infecting corneal cells. Unexpectedly, the receptor is constituted by one or more glycoproteins containing the GD1a glycan motif rather than the ganglioside itself, as shown by binding, infection and flow cytometry experiments. Molecular modeling, nuclear magnetic resonance and X-ray crystallography reveal that the two terminal sialic acids dock into two of three previously established sialic acid–binding sites in the trimeric Ad37 knob. Surface plasmon resonance analysis shows that the knob–GD1a glycan interaction has high affinity. Our findings therefore form a basis for the design and development of sialic acid–containing antiviral drugs for topical treatment of EKC.

Your institute does not have access to this article

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.

$32.00

All prices are NET prices.

Figure 1: Glycan array of Ad37 knob interactions with 260 different glycans.
Figure 2: Ad37 binding to the corneal epithelial cell surface depends on fiber knob binding to molecules containing the GD1a glycan.
Figure 3: X-ray crystallography and functional analysis of the Ad37 knob-GD1a glycan complex.
Figure 4: Surface plasmon resonance analysis of GD1a glycan binding to immobilized Ad37 knob protein.

References

  1. Wold, W.S.M. & Horwitz, M.S. Adenoviruses. in Fields Virology, Vol. 2 (eds. Knipe, D.M. & Howley, P.M.) 2395–2436. (Lippincott Williams & Wilkins, Philadelphia, 2007).

  2. Ford, E., Nelson, K.E. & Warren, D. Epidemiology of epidemic keratoconjunctivitis. Epidemiol. Rev. 9, 244–261 (1987).

    Google Scholar 

  3. Arnberg, N. Adenovirus receptors, implications for tropism, treatment and targeting. Rev. Med. Virol. 19, 165–178 (2009).

    Google Scholar 

  4. Arnberg, N., Edlund, K., Kidd, A.H. & Wadell, G. Adenovirus type 37 uses sialic acid as a cellular receptor. J. Virol. 74, 42–48 (2000).

    Google Scholar 

  5. Gordon, Y.J., Aoki, K. & Kinchington, P.R. Adenovirus keratoconjunctivitis. in Ocular Infection and Immunity (eds. Pepose, J.S., Holland, G.N. & Wilhelmus, K.R.) 877–894. (Mosby, St. Louis, 1996).

  6. Kinchington, P.R., Romanowski, E.G. & Gordon, Y.J. Prospects for adenovirus antivirals. J. Antimicrob. Chemother. 55, 424–429 (2005).

    Google Scholar 

  7. Arnberg, N. et al. Adenovirus type 37 binds to cell surface sialic acid through a charge-dependent interaction. Virology 302, 33–43 (2002).

    Google Scholar 

  8. Arnberg, N., Pring-Akerblom, P. & Wadell, G. Adenovirus type 37 uses sialic acid as a cellular receptor on Chang C cells. J. Virol. 76, 8834–8841 (2002).

    Google Scholar 

  9. Wu, E. et al. Membrane cofactor protein is a receptor for adenoviruses associated with epidemic keratoconjunctivitis. J. Virol. 78, 3897–3905 (2004).

    Google Scholar 

  10. Cashman, S.M., Morris, D.J. & Kumar-Singh, R. Adenovirus type 5 pseudotyped with adenovirus type 37 fiber uses sialic acid as a cellular receptor. Virology 324, 129–139 (2004).

    Google Scholar 

  11. Lecollinet, S. et al. Improved gene delivery to intestinal mucosa by adenoviral vectors bearing subgroup B and D fibers. J. Virol. 80, 2747–2759 (2006).

    Google Scholar 

  12. Thirion, C. et al. Adenovirus vectors based on human adenovirus type 19a have high potential for human muscle-directed gene therapy. Hum. Gene Ther. 17, 193–205 (2006).

    Google Scholar 

  13. Arnberg, N., Kidd, A.H., Edlund, K., Olfat, F. & Wadell, G. Initial interactions of subgenus D adenoviruses with A549 cellular receptors: sialic acid versus αv integrins. J. Virol. 74, 7691–7693 (2000).

    Google Scholar 

  14. Rinaldi, S. et al. Analysis of lectin binding to glycolipid complexes using combinatorial glycoarrays. Glycobiology 19, 789–796 (2009).

    Google Scholar 

  15. Mayer, M. & Meyer, B. Characterization of ligand binding by saturation transfer difference NMR spectroscopy. Angew. Chem. Int. Ed. 38, 1784–1788 (1999).

    Google Scholar 

  16. Burmeister, W.P., Guilligay, D., Cusack, S., Wadell, G. & Arnberg, N. Crystal structure of species D adenovirus fiber knobs and their sialic acid binding sites. J. Virol. 78, 7727–7736 (2004).

    Google Scholar 

  17. Bewley, M.C., Springer, K., Zhang, Y.B., Freimuth, P. & Flanagan, J.M. Structural analysis of the mechanism of adenovirus binding to its human cellular receptor, CAR. Science 286, 1579–1583 (1999).

    Google Scholar 

  18. Persson, B.D. et al. Adenovirus type 11 binding alters the conformation of its receptor CD46. Nat. Struct. Mol. Biol. 14, 164–166 (2007).

    Google Scholar 

  19. Kirby, I. et al. Identification of contact residues and definition of the CAR-binding site of adenovirus type 5 fiber protein. J. Virol. 74, 2804–2813 (2000).

    Google Scholar 

  20. Sauter, N.K. et al. Hemagglutinins from two influenza virus variants bind to sialic acid derivatives with millimolar dissociation constants: a 500-MHz proton nuclear magnetic resonance study. Biochemistry 28, 8388–8396 (1989).

    Google Scholar 

  21. Arnberg, N., Mei, Y. & Wadell, G. Fiber genes of adenoviruses with tropism for the eye and the genital tract. Virology 227, 239–244 (1997).

    Google Scholar 

  22. Lenaerts, L., De Clercq, E. & Naesens, L. Clinical features and treatment of adenovirus infections. Rev. Med. Virol. 18, 357–374 (2008).

    Google Scholar 

  23. von Itzstein, M. et al. Rational design of potent sialidase-based inhibitors of influenza virus replication. Nature 363, 418–423 (1993).

    Google Scholar 

  24. Kim, C.U. et al. Influenza neuraminidase inhibitors possessing a novel hydrophobic interaction in the enzyme active site: design, synthesis, and structural analysis of carbocyclic sialic acid analogues with potent anti-influenza activity. J. Am. Chem. Soc. 119, 681–690 (1997).

    Google Scholar 

  25. Johansson, S.M. et al. Multivalent sialic acid conjugates inhibit adenovirus type 37 from binding to and infecting human corneal epithelial cells. Antiviral Res. 73, 92–100 (2007).

    Google Scholar 

  26. Araki-Sasaki, K. et al. An SV40-immortalized human corneal epithelial cell line and its characterization. Invest. Ophthalmol. Vis. Sci. 36, 614–621 (1995).

    Google Scholar 

  27. Tsai, B. et al. Gangliosides are receptors for murine polyoma virus and SV40. EMBO J. 22, 4346–4355 (2003).

    Google Scholar 

  28. Boffey, J. et al. Characterisation of the immunoglobulin variable region gene usage encoding the murine anti-ganglioside antibody repertoire. J. Neuroimmunol. 165, 92–103 (2005).

    Google Scholar 

  29. Kabsch, W. Automatic processing of rotation diffraction data from crystals of initially unknown symmetry and cell constants. J. Appl. Crystallogr. 26, 795–800 (1993).

    Google Scholar 

  30. McCoy, A.J. et al. Phaser crystallographic software. J. Appl. Crystallogr. 40, 658–674 (2007).

    Google Scholar 

  31. Collaborative Computational Project. The CCP4 suite: programs for protein crystallography. Acta Crystallogr. D Biol. Crystallogr. 50, 760–763 (1994).

  32. Murshudov, G.N., Vagin, A.A. & Dodson, E.J. Refinement of Macromolecular Structures by the Maximum-Likelihood Method. Acta Crystallogr. D Biol. Crystallogr. 53, 240–255 (1997).

    Google Scholar 

  33. Emsley, P. & Cowtan, K. Coot: model building tools for molecular graphics. Acta Crystallogr. D Biol. Crystallogr. 60, 2126–2132 (2004).

    Google Scholar 

  34. Adams, P.D. et al. PHENIX: a comprehensive Python-based system for macromolecular structure solution. Acta Crystallogr. D Biol. Crystallogr. 66, 213–221 (2010).

    Google Scholar 

Download references

Acknowledgements

We highly appreciate the support from F. Lindh and S. Spjut regarding the purification of GD1a glycan; F. Lindh (Isosep) provided GD1a gangliosides. We also thank A. Carlsson (Medigelium) for providing GD1a-containing liposomes; D. Guilligay (EMBL) for providing the Ad37 knob gene cloned into pPROEX Htb plasmid (Life Technologies)16; R.L. Schnaar (The Johns Hopkins University School of Medicine) for providing P4 compounds; and K. Lindman, M. Hägg and F. Jamshidi for technical support. HCE cells were provided by K. Araki-Sasaki (Kinki Central Hospital). GM3 and GD2 glycans were provided by the Consortium for Functional Glycomics. We also highly appreciate the resources provided by the Consortium for Functional Glycomics (funded by National Institute of General Medical Sciences grant no. GM62116) Core D and H, as well as related technical support from O. Blixt and N. Reza. We are grateful to the Berlin Electron Storage Ring Society for Synchrotron Radiation (BESSY) for beamtime and beamline support. This project was supported by the Swedish Research Council (grants no. 2007-3402 (N.A.); 2009-3859 (N.A.) and 11612 (J.Å. via M.E. Breimer)), the Swedish Foundation for Strategic Research (grant no. F06-0011 to N.A.), the Swedish Society of Medicine (grant no. 97031 to N.A.), the Estonian Science Foundation (grant 8300 to A.L.), the Collaborative Research Center SFB-685 (T.S.), a student fellowship from the University of Tübingen (J.B.) and The Wellcome Trust (S.R. and H.J.W.).

Author information

Authors and Affiliations

Authors

Contributions

E.C.N. and R.J.S. contributed equally to design and conduction of binding, infection and flow cytometry experiments; S.M.C.J. produced GD1a glycan and performed NMR studies with M.H.; J.B. and T.S. carried out X-ray crystallography studies; S.R. and H.J.W. performed combinatorial glycolipid glycoarray; J.Å. conducted molecular modeling; F.P.D. carried out immunohistochemistry analysis; and A.L. performed SPR experiments. L.F. and T.L.E. conducted two-dimensional gel electrophoresis and blotting experiments, and L.F. did statistical calculations. T.S., M.H., F.P.D., A.L., J.Å., H.J.W., L.F., J.B. and N.A. discussed and wrote the manuscript, and T.S. and N.A. supervised the project.

Corresponding author

Correspondence to Niklas Arnberg.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–6 and Supplementary Tables 2–5 (PDF 4855 kb)

Supplementary Table 1

Summary of glycan array data (XLS 119 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Nilsson, E., Storm, R., Bauer, J. et al. The GD1a glycan is a cellular receptor for adenoviruses causing epidemic keratoconjunctivitis. Nat Med 17, 105–109 (2011). https://doi.org/10.1038/nm.2267

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nm.2267

Further reading

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing