1887

Abstract

Influenza virus infects host cells through membrane fusion, a process mediated by the low pH-induced conformational change of the viral surface glycoprotein haemagglutinin (HA). We determined the structures and biochemical properties of the HA proteins from A/Korea/01/2009 (KR01), a 2009 pandemic strain, and A/Thailand/CU44/2006 (CU44), a seasonal strain. The crystal structure of KR01 HA revealed a V-shaped head-to-head arrangement, which is not seen in other HA proteins including CU44 HA. We isolated a broadly neutralizing H1-specific monoclonal antibody GC0757. The KR01 HA-Fab0757 complex structure also exhibited a head-to-head arrangement of HA. Both native and Fab complex structures reveal a different spatial orientation of HA1 relative to HA2, indicating that HA is flexible and dynamic at neutral pH. Further, the KR01 HA exhibited significantly lower protein stability and increased susceptibility to proteolytic cleavage compared with other HAs. Our structures provide important insights into the conformational flexibility of HA.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.051136-0
2013-08-01
2019-09-19
Loading full text...

Full text loading...

/deliver/fulltext/jgv/94/8/1712.html?itemId=/content/journal/jgv/10.1099/vir.0.051136-0&mimeType=html&fmt=ahah

References

  1. Adams P. D. , Afonine P. V. , Bunkóczi G. , Chen V. B. , Davis I. W. , Echols N. , Headd J. J. , Hung L. W. , Kapral G. J. . & other authors ( 2010; ). PHENIX: a comprehensive Python-based system for macromolecular structure solution. . Acta Crystallogr D Biol Crystallogr 66:, 213–221. [CrossRef] [PubMed]
    [Google Scholar]
  2. Bhardwaj A. , Walker-Kopp N. , Wilkens S. , Cingolani G. . ( 2008; ). Foldon-guided self-assembly of ultra-stable protein fibers. . Protein Sci 17:, 1475–1485. [CrossRef] [PubMed]
    [Google Scholar]
  3. Böttcher E. , Matrosovich T. , Beyerle M. , Klenk H. D. , Garten W. , Matrosovich M. . ( 2006; ). Proteolytic activation of influenza viruses by serine proteases TMPRSS2 and HAT from human airway epithelium. . J Virol 80:, 9896–9898. [CrossRef] [PubMed]
    [Google Scholar]
  4. Brünger A. T. . ( 1992; ). Free R value: a novel statistical quantity for assessing the accuracy of crystal structures. . Nature 355:, 472–475. [CrossRef] [PubMed]
    [Google Scholar]
  5. Bullough P. A. , Hughson F. M. , Skehel J. J. , Wiley D. C. . ( 1994; ). Structure of influenza haemagglutinin at the pH of membrane fusion. . Nature 371:, 37–43. [CrossRef] [PubMed]
    [Google Scholar]
  6. Carr C. M. , Kim P. S. . ( 1993; ). A spring-loaded mechanism for the conformational change of influenza hemagglutinin. . Cell 73:, 823–832. [CrossRef] [PubMed]
    [Google Scholar]
  7. Carr C. M. , Chaudhry C. , Kim P. S. . ( 1997; ). Influenza hemagglutinin is spring-loaded by a metastable native conformation. . Proc Natl Acad Sci U S A 94:, 14306–14313. [CrossRef] [PubMed]
    [Google Scholar]
  8. Chaipan C. , Kobasa D. , Bertram S. , Glowacka I. , Steffen I. , Tsegaye T. S. , Takeda M. , Bugge T. H. , Kim S. . & other authors ( 2009; ). Proteolytic activation of the 1918 influenza virus hemagglutinin. . J Virol 83:, 3200–3211. [CrossRef] [PubMed]
    [Google Scholar]
  9. Conenello G. M. , Zamarin D. , Perrone L. A. , Tumpey T. , Palese P. . ( 2007; ). A single mutation in the PB1-F2 of H5N1 (HK/97) and 1918 influenza A viruses contributes to increased virulence. . PLoS Pathog 3:, 1414–1421. [CrossRef] [PubMed]
    [Google Scholar]
  10. Danieli T. , Pelletier S. L. , Henis Y. I. , White J. M. . ( 1996; ). Membrane fusion mediated by the influenza virus hemagglutinin requires the concerted action of at least three hemagglutinin trimers. . J Cell Biol 133:, 559–569. [CrossRef] [PubMed]
    [Google Scholar]
  11. DuBois R. M. , Aguilar-Yañez J. M. , Mendoza-Ochoa G. I. , Oropeza-Almazán Y. , Schultz-Cherry S. , Alvarez M. M. , White S. W. , Russell C. J. . ( 2011; ). The receptor-binding domain of influenza virus hemagglutinin produced in Escherichia coli folds into its native, immunogenic structure. . J Virol 85:, 865–872. [CrossRef] [PubMed]
    [Google Scholar]
  12. Emsley P. , Cowtan K. . ( 2004; ). Coot: model-building tools for molecular graphics. . Acta Crystallogr D Biol Crystallogr 60:, 2126–2132. [CrossRef] [PubMed]
    [Google Scholar]
  13. Feshchenko E. , Rhodes D. G. , Felberbaum R. F. , McPherson C. , Rininger J. A. , Post P. , Cox M. M. J. . ( 2012; ). Pandemic influenza vaccine: characterization of A/California/07/2009 (H1N1) recombinant hemagglutinin protein and insights into H1N1 antigen stability. . BMC Biotechnol 12:, 77. [CrossRef] [PubMed]
    [Google Scholar]
  14. Floyd D. L. , Ragains J. R. , Skehel J. J. , Harrison S. C. , van Oijen A. M. . ( 2008; ). Single-particle kinetics of influenza virus membrane fusion. . Proc Natl Acad Sci U S A 105:, 15382–15387. [CrossRef] [PubMed]
    [Google Scholar]
  15. Fontana J. , Cardone G. , Heymann J. B. , Winkler D. C. , Steven A. C. . ( 2012; ). Structural changes in influenza virus at low pH characterized by cryo-electron tomography. . J Virol 86:, 2919–2929. [CrossRef] [PubMed]
    [Google Scholar]
  16. Fraser C. , Donnelly C. A. , Cauchemez S. , Hanage W. P. , Van Kerkhove M. D. , Hollingsworth T. D. , Griffin J. , Baggaley R. F. , Jenkins H. E. . & other authors ( 2009; ). Pandemic potential of a strain of influenza A (H1N1): early findings. . Science 324:, 1557–1561. [CrossRef] [PubMed]
    [Google Scholar]
  17. Gamblin S. J. , Haire L. F. , Russell R. J. , Stevens D. J. , Xiao B. , Ha Y. , Vasisht N. , Steinhauer D. A. , Daniels R. S. . & other authors ( 2004; ). The structure and receptor binding properties of the 1918 influenza hemagglutinin. . Science 303:, 1838–1842. [CrossRef] [PubMed]
    [Google Scholar]
  18. Garten R. J. , Davis C. T. , Russell C. A. , Shu B. , Lindstrom S. , Balish A. , Sessions W. M. , Xu X. , Skepner E. . & other authors ( 2009; ). Antigenic and genetic characteristics of swine-origin 2009 A(H1N1) influenza viruses circulating in humans. . Science 325:, 197–201. [CrossRef] [PubMed]
    [Google Scholar]
  19. Harrison S. C. . ( 2008; ). Viral membrane fusion. . Nat Struct Mol Biol 15:, 690–698. [CrossRef] [PubMed]
    [Google Scholar]
  20. Huang S. S. , Banner D. , Fang Y. , Ng D. C. , Kanagasabai T. , Kelvin D. J. , Kelvin A. A. . ( 2011; ). Comparative analyses of pandemic H1N1 and seasonal H1N1, H3N2, and influenza B infections depict distinct clinical pictures in ferrets. . PLoS ONE 6:, e27512. [CrossRef] [PubMed]
    [Google Scholar]
  21. Itoh Y. , Shinya K. , Kiso M. , Watanabe T. , Sakoda Y. , Hatta M. , Muramoto Y. , Tamura D. , Sakai-Tagawa Y. . & other authors ( 2009; ). In vitro and in vivo characterization of new swine-origin H1N1 influenza viruses. . Nature 460:, 1021–1025.[PubMed]
    [Google Scholar]
  22. Kendal A. P. , Skehel J. J. , Pereira M. S. . ( 1982; ). Concepts and Procedures for Laboratory Based Influenza Surveillance. Atlanta, GA:: Centers for Disease Control;.
    [Google Scholar]
  23. Khurana S. , Verma S. , Verma N. , Crevar C. J. , Carter D. M. , Manischewitz J. , King L. R. , Ross T. M. , Golding H. . ( 2010; ). Properly folded bacterially expressed H1N1 hemagglutinin globular head and ectodomain vaccines protect ferrets against H1N1 pandemic influenza virus. . PLoS ONE 5:, e11548. [CrossRef] [PubMed]
    [Google Scholar]
  24. Laskowski R. A. , MacArthus M. W. , Moss D. S. , Thornton J. M. . ( 1993; ). PROCHECK: a program to check the stereochemical quality of protein structures. . J Appl Crystallogr 26:, 283–291. [CrossRef]
    [Google Scholar]
  25. Lazarowitz S. G. , Choppin P. W. . ( 1975; ). Enhancement of the infectivity of influenza A and B viruses by proteolytic cleavage of the hemagglutinin polypeptide. . Virology 68:, 440–454. [CrossRef] [PubMed]
    [Google Scholar]
  26. Lee J. H. , Goulian M. , Boder E. T. . ( 2006; ). Autocatalytic activation of influenza hemagglutinin. . J Mol Biol 364:, 275–282. [CrossRef] [PubMed]
    [Google Scholar]
  27. Maines T. R. , Jayaraman A. , Belser J. A. , Wadford D. A. , Pappas C. , Zeng H. , Gustin K. M. , Pearce M. B. , Viswanathan K. . & other authors ( 2009; ). Transmission and pathogenesis of swine-origin 2009 A(H1N1) influenza viruses in ferrets and mice. . Science 325:, 484–487.[PubMed]
    [Google Scholar]
  28. Markovic I. , Leikina E. , Zhukovsky M. , Zimmerberg J. , Chernomordik L. V. . ( 2001; ). Synchronized activation and refolding of influenza hemagglutinin in multimeric fusion machines. . J Cell Biol 155:, 833–844. [CrossRef] [PubMed]
    [Google Scholar]
  29. Munster V. J. , de Wit E. , van den Brand J. M. , Herfst S. , Schrauwen E. J. , Bestebroer T. M. , van de Vijver D. , Boucher C. A. , Koopmans M. . & other authors ( 2009; ). Pathogenesis and transmission of swine-origin 2009 A(H1N1) influenza virus in ferrets. . Science 325:, 481–483.[PubMed]
    [Google Scholar]
  30. Murshudov G. N. , Vagin A. A. , Dodson E. J. . ( 1997; ). Refinement of macromolecular structures by the maximum-likelihood method. . Acta Crystallogr D Biol Crystallogr 53:, 240–255. [CrossRef] [PubMed]
    [Google Scholar]
  31. Otwinowski Z. , Minor W. . ( 1997; ). Processing of X-ray diffraction data collected in oscillation mode. . In Methods in Enzymology, Volume 276: Macromolecular Crystallography, Part A, pp. 307–326. Edited by Carter C. W. Jr. , Sweet R. M. . . San Diego:: Academic Press;. [CrossRef] [PubMed]
    [Google Scholar]
  32. Safronetz D. , Rockx B. , Feldmann F. , Belisle S. E. , Palermo R. E. , Brining D. , Gardner D. , Proll S. C. , Marzi A. . & other authors ( 2011; ). Pandemic swine-origin H1N1 influenza A virus isolates show heterogeneous virulence in macaques. . J Virol 85:, 1214–1223. [CrossRef] [PubMed]
    [Google Scholar]
  33. Salomon R. , Webster R. G. . ( 2009; ). The influenza virus enigma. . Cell 136:, 402–410. [CrossRef] [PubMed]
    [Google Scholar]
  34. Shaw N. , Zhao M. , Cheng C. , Xu H. , Saarikettu J. , Li Y. , Da Y. , Yao Z. , Silvennoinen O. . & other authors ( 2007; ). The multifunctional human p100 protein ‘hooks’ methylated ligands. . Nat Struct Mol Biol 14:, 779–784. [CrossRef] [PubMed]
    [Google Scholar]
  35. Skehel J. J. , Wiley D. C. . ( 2000; ). Receptor binding and membrane fusion in virus entry: the influenza hemagglutinin. . Annu Rev Biochem 69:, 531–569. [CrossRef] [PubMed]
    [Google Scholar]
  36. Steel J. , Lowen A. C. , Mubareka S. , Palese P. . ( 2009; ). Transmission of influenza virus in a mammalian host is increased by PB2 amino acids 627K or 627E/701N. . PLoS Pathog 5:, e1000252. [CrossRef] [PubMed]
    [Google Scholar]
  37. Stevens J. , Corper A. L. , Basler C. F. , Taubenberger J. K. , Palese P. , Wilson I. A. . ( 2004; ). Structure of the uncleaved human H1 hemagglutinin from the extinct 1918 influenza virus. . Science 303:, 1866–1870. [CrossRef] [PubMed]
    [Google Scholar]
  38. The CCP4. ( 1994; ). The CCP4 suite: programs for protein crystallography. . Acta Crystallogr D Biol Crystallogr 50:, 760–763.[CrossRef]
    [Google Scholar]
  39. Wang Q. , Cheng F. , Lu M. , Tian X. , Ma J. . ( 2008; ). Crystal structure of unliganded influenza B virus hemagglutinin. . J Virol 82:, 3011–3020. [CrossRef] [PubMed]
    [Google Scholar]
  40. Wilson I. A. , Skehel J. J. , Wiley D. C. . ( 1981; ). Structure of the haemagglutinin membrane glycoprotein of influenza virus at 3 Å resolution. . Nature 289:, 366–373. [CrossRef] [PubMed]
    [Google Scholar]
  41. Xu R. , Wilson I. A. . ( 2011; ). Structural characterization of an early fusion intermediate of influenza virus hemagglutinin. . J Virol 85:, 5172–5182. [CrossRef] [PubMed]
    [Google Scholar]
  42. Xu R. , Ekiert D. C. , Krause J. C. , Hai R. , Crowe J. E. Jr , Wilson I. A. . ( 2010; ). Structural basis of preexisting immunity to the 2009 H1N1 pandemic influenza virus. . Science 328:, 357–360. [CrossRef] [PubMed]
    [Google Scholar]
  43. Yang, H., Carney, P. & Stevens, J. (2010). Structure and receptor binding properties of a pandemic H1N1 virus hemagglutinin. PLoS Curr Influenza RRN1152.
  44. Zambon M. C. . ( 2001; ). The pathogenesis of influenza in humans. . Rev Med Virol 11:, 227–241. [CrossRef] [PubMed]
    [Google Scholar]
  45. Zhang W. , Qi J. , Shi Y. , Li Q. , Gao F. , Sun Y. , Lu X. , Lu Q. , Vavricka C. J. . & other authors ( 2010; ). Crystal structure of the swine-origin A (H1N1)-2009 influenza A virus hemagglutinin (HA) reveals similar antigenicity to that of the 1918 pandemic virus. . Protein Cell 1:, 459–467. [CrossRef] [PubMed]
    [Google Scholar]
  46. Zimmer S. M. , Burke D. S. . ( 2009; ). Historical perspective–Emergence of influenza A (H1N1) viruses. . N Engl J Med 361:, 279–285. [CrossRef] [PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.051136-0
Loading
/content/journal/jgv/10.1099/vir.0.051136-0
Loading

Data & Media loading...

Supplementary material 

PDF

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error