Structure of a pertussis toxin–sugar complex as a model for receptor binding

Abstract

Pertussis toxin is an exotoxin from the bacterium Bordetella pertussis which is important the pathogenesis of whooping cough and the generation of a protective immune response. The diverse biological activities of the toxin depend on its ability to recognize carbohydrate-containing receptors on a wide variety of eukaryotic cells. We present here the crystal structure of pertussis toxin complexed with a soluble oligosaccharide from transferrin. Binding sites for the terminal sialic acid–galactose moiety are revealed on both subunits S2 and S3 of the B-oligomer. Identification of amino acid residues involved in receptor binding will improve the design of genetically inactivated toxins for use in new acellular whooping cough vaccines

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References

  1. 1

    Munoz, J.J., Arai, H., Bergman, R.K. & Sadowski, P.L. Biological activities of crystalline pertussigen from Bordetella pertussis. Infect. Immun. 33, 820–826 (1981).

  2. 2

    Armstrong, G.D., Howard, L.A. & Peppler, M.S. Use of glycosyltransferases to restore pertussis toxin receptor activity to asialogalactofetuin. J. biol. Chem. 263, 8677–8684 (1988).

  3. 3

    Brennan, M.J., David, J.L., Kenimer, J.G. & Manclark, C.R. Lectin-like binding of pertussis toxin to a 165-kilodalton Chinese hamster ovary cell glycoprotein. J. biol. Chem. 263, 4895–4899 (1988).

  4. 4

    Witvliet, M.H., Burns, D.L., Brennan, M.J., Poolman, J.T. & Manclark, C.R. Binding of pertussis toxin to eukaryotic cells and glycoproteins. Infect. Immun. 57, 3324–3330 (1989).

  5. 5

    Clark, C.G. & Armstrong, G.D. Lymphocyte receptors for pertussis toxin. Infect. Immun. 58, 3840–3846 (1990).

  6. 6

    Heerze, L.D., Chong, P.C.S. & Armstrong, G.D. Investigation of the lectin-like binding domains in pertussis toxin using synthetic peptide sequences. J. biol. Chem. 267, 25810–25815 (1992).

  7. 7

    Saukkonen, K., Burnette, W.N., Mar, V.L., Masure, H.R. & Tuomanen, E.I. Pertussis toxin has eukaryotic-like carbohydrate recognition domains. Proc. natn. Acad. Sci. U.S.A. 89, 118–122 (1992).

  8. 8

    van't Wout, J., et al. Role of carbohydrate recognition domains of pertussis toxin in adherence of Bordetella pertussis to human macrophages. Infect. Immun. 60, 3303–3308 (1992).

  9. 9

    Stein, P.E. et al. The crystal structure of pertussis toxin. Structure 2, 45–57 (1994).

  10. 10

    Weis, W.I., Kahn, R., Fourme, R., Drickamer, K. & Hendrickson, W.A. Structure of the calcium-dependent lectin domain from a rat mannose-binding protein determined by MAD phasing. Science 254, 1608–1615 (1991).

  11. 11

    Graves, B.J. et al. Insight into E-selectin/ligand interaction from the crystal structure and mutagenesis of the lec/EGF domains. Nature 367, 532–538 (1994).

  12. 12

    Murzin, A.G. OB (oligonucleotide/oligosaccharide binding)-fold: common structural and functional solution for non-homologous sequences. EMBO J. 12, 861–867 (1993).

  13. 13

    Spik, G. et al. Studies on glycoconjugates. LXIV. Complete structure of two carbohydrate units of human serotransferrin. FEBS Lett. 50, 296–299 (1975).

  14. 14

    Breg, J., Kroon-Batenburg, L.M.J., Strecker, G., Montreuil, J. & Vliegenthart, J.F.G. Conformational analysis of the sialylα(2→3/6)N-acetyllactosamine structural element occurring in glycoproteins by two-dimensional NOE 1H-NMR spectroscopy in combination with energy calculations by hard-sphere exo-anomeric and molecular mechanics force-field with hydrogen-bonding potential. Eur. J. Biochem. 178, 727–739 (1989).

  15. 15

    Spangler, B.D., Heerze, L.D., Clark, C.G. & Armstrong, G.D. Hydrophobic binding of pertussis toxin is enhanced by oligosaccharide receptors. Arch. Biochem. Biophys. 305, 153–158 (1993).

  16. 16

    Shigeta, R., Forest, K., Yan, L., Kahne, D. & Schutt, C.E. Isomorphous binding of mercury-substituted thiosaccharides to pertussis toxin crystals yields crystallographic phases. Acta crystallogr. D50, 71–74 (1994).

  17. 17

    Merritt, E.A. et al. Crystal structure of cholera toxin B-pentamer bound to receptor GM1 pentasaccharide. Prot. Sci. 3, 166–175 (1994).

  18. 18

    Sixma, T.K. et al. Lactose binding to heat-labile enterotoxin revealed by X-ray crystallography. Nature 355, 561–564 (1992).

  19. 19

    Armstrong, G.D. & Peppler, M.S. Maintenance of biological activity of pertussis toxin radioiodinated while bound to fetuin-agarose. Infect. Immun. 55, 1294–1299 (1987).

  20. 20

    Lobet, Y. et al. Site-specific alterations in the B-oligomer that affect receptor-binding activities and mitogenicity of pertussis toxin. J. exp. Med. 177, 79–87 (1993).

  21. 21

    Loosmore, S. et al. Characterization of pertussis toxin analogs containing mutations in the B-oligomer subunits. Infect. Immun. 61, 2316–2324 (1993).

  22. 22

    Weis, W.I., Drickamer, K. & Hendickson, W.A. Structure of a C-type mannose-binding protein complexed with an oligosaccharide. Nature 360, 127–134 (1992).

  23. 23

    Sato, Y., Sato, H., Tiru, M. & Brown, F. Pertussis: evaluation and research on acellular pertussis vaccines (Karger, Basel, 1991).

  24. 24

    Hewlett, E.L. & Cowell, J.L. Evaluation of the mouse model for study of encehalopathy in pertussis vaccine recipients. Infect. Immun. 57, 661–663 (1981).

  25. 25

    Burnette, W.N. et al. Pertussis toxin S1 mutant with reduced enzyme activity and a conserved protective epitope. Science 242, 72–74 (1988).

  26. 26

    Pizza, M., Bartoloni, A., Prugnola, A., Silvestri, S. & Rappuoli, R. Subunit S1 of pertussis toxin: mapping of the regions essential for ADP-ribosyltransferase activity. Proc. natn. Acad. Sci. U.S.A. 85, 7521–7525 (1988).

  27. 27

    Loosmore, S.M. et al. Engineering of genetically detoxified pertussis toxin analogs for development of a recombinant whooping cough vaccine. Infect. Immun. 58, 3653–3662 (1990).

  28. 28

    Nencioni, L. et al. Characterization of genetically inactivated pertussis toxin mutants: candidates for a new vaccine against whooping cough. Infect. Immun. 58, 1308–1315 (1990).

  29. 29

    Kimura, A., Mountzouros, K.T., Schad, P.A., Cieplak, W. & Cowell, J.L. Pertussis toxin analog with reduced enzymatic and biological activities is a protective antigen. Infect. Immun. 58, 3337–3347 (1990).

  30. 30

    Sakabe, N. A focusing Weissenberg camera with multi-layer-line screens for macromolecular crystallography. J. appl. Crystallogr. 16, 542–547 (1983).

  31. 31

    Higashi, T. The processing of diffraction data taken on a screenless Weissenberg camera for macromolecular crystallography. J. appl. Crystallogr. 22, 9–18 (1989).

  32. 32

    Brünger, A.T., Kuriyan, J. & Karplus, M. Crystallographic R-factor refinement by molecular graphics. Science 235, 458–460 (1987).

  33. 33

    Read, R.J. Improved Fourier coefficients for maps using phases from partial structures with errors. Acta crystallogr. A42, 140–149 (1986).

  34. 34

    Jones, T.A., Zou, J.-Y., Cowan, S.W. & Kjeldgaard, M. Improved methods for building protein models in electron density maps and the location of errors in these models. Acta crystallogr. A47, 110–119 (1991).

  35. 35

    Weis, W.I., Brünger, A.T., Skehel, J.J. & Wiley, D.C. Refinement of the influenza virus haemagglutinin by simulated annealing. J. molec. Biol. 212, 737–761 (1990).

  36. 36

    Kraulis, P. MOLSCRIPT: a program to produce both detailed and schematic plots of proteins. J. appl. Crystallogr. 24, 946–950 (1991).

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Stein, P., Boodhoo, A., Armstrong, G. et al. Structure of a pertussis toxin–sugar complex as a model for receptor binding. Nat Struct Mol Biol 1, 591–596 (1994). https://doi.org/10.1038/nsb0994-591

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