1887

Abstract

SUMMARY

Haemagglutinin prepared from influenza virus A/Memphis/1/71 by bromelain digestion was centrifuged through continuous sucrose gradients buffered at pH 7.4 or pH 4.9. From these gradients were isolated two forms of the protein which displayed different equilibrium sedimentation properties. One species behaved as a molecule with a mol. wt. of 190000, the other with a mol. wt. of 70000. These results are consistent with the separation of trimeric and monomeric haemagglutinin. A comparison of their antigenic properties, using monoclonal antibodies raised against intact virus, showed that major antigenic differences occur between the two forms of haemagglutinin. None of the monoclonal antibodies reacted with haemagglutinin denatured by reduction and alkylation.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-66-8-1687
1985-08-01
2024-04-18
Loading full text...

Full text loading...

/deliver/fulltext/jgv/66/8/JV0660081687.html?itemId=/content/journal/jgv/10.1099/0022-1317-66-8-1687&mimeType=html&fmt=ahah

References

  1. Bartel A. H., Campbell D. H. 1959; Some immunochemical differences between associated and dissociated hemocyanin. Archives of Biochemistry and Biophysics 82:232–34
    [Google Scholar]
  2. Bothwell M. A., Howlett O. J., Schachman H. K. 1978; A sedimentation equilibrium method for determining molecular weights of proteins with a tabletop high speed air turbine centrifuge. Journal of Biological Chemistry 253:2073–2077
    [Google Scholar]
  3. Brand C. M., Skehel J. J. 1972; Crystalline antigens from the influenza virus envelope. Nature New Biology 238:145–147
    [Google Scholar]
  4. Brown L. E., Dopheide T. A. A., Ward C. W., White D. O., Jackson D. C. 1980; Antigenic determinants of influenza virus hemagglutinin. V. Antigenicity of the HA2 chain. Journal of Immunology 125:1583–1588
    [Google Scholar]
  5. Daniels R. S., Douglas A. R., Skehel J. J., Wiley D. C. 1983; Analyses of the antigenicity of influenza haemagglutinin at the pH optimum for virus-mediated membrane fusion. Journal of General Virology 64:1657–1662
    [Google Scholar]
  6. Ey L., Prowse S. J., Jenkin C. R. 1978; Isolation of pure IgG1, IgG2a and IgG2b immunoglobulins from mouse serum using protein A-Sepharose. Immunochemistry 15:429–436
    [Google Scholar]
  7. Greenwood F. C., Hunter W. M., Glover J. S. 1963; The preparation of 131I-labelled human growth hormone of high specific radioactivity. Biochemical Journal 89:114–123
    [Google Scholar]
  8. Hahn E., Timpl R. 1973; Involvement of more than a single polypeptide chain in the helical antigenic determinants of collagen. European Journal of Immunology 3:442–446
    [Google Scholar]
  9. Jackson D. C., Nestorowicz A. 1985; Antigenic determinants of influenza virus hemagglutinin. XI. Evidence for a hinge bending mechanism. Virology (in press)
    [Google Scholar]
  10. Jackson D. C., Brown L. E., White D. O., Dopheide T. A. A., Ward C. W. 1979; Antigenic determinants of influenza virus hemagglutinin. IV. Immunogenicity of fragments isolated from the hemagglutinin of A/Memphis/72. Journal of Immunology 123:2610–2617
    [Google Scholar]
  11. Jackson D. C., Murray J. M., Brown L. E., White D. O. 1981; Immunochemical properties of influenza virus hemagglutinin and its fragments. In Genetic Variation Among Influenza Viruses pp 355–372 Edited by Nayak D. P. New York: Academic Press;
    [Google Scholar]
  12. Jackson D. C., Murray J. M., Anders E. M., White D. O., Webster R. G., Brown L. E. 1983; Expression of a unique antigenic determinant of influenza virus hemagglutinin at pH 5. In The Origin of Pandemic Influenza Viruses p 29 Edited by Laver W. G. New York: Elsevier;
    [Google Scholar]
  13. Laver W. G. 1969; Purification of influenza viruses. In Fundamental Techniques in Virology p 82 Edited by Habel K., Salzman N. P. New York: Academic Press;
    [Google Scholar]
  14. Laver W. G., Air G. M., Webster R. G., Gerhard W., Ward C. W., Dopheide T. A. A. 1979; Antigenic drift in type A influenza viruses: sequence differences in the hemagglutinin of Hong Kong (H3N2) variants selected with monoclonal hybridoma antibodies. Virology 98:226–237
    [Google Scholar]
  15. Newton S. E., Air G. M., Webster R. G., Laver W. G. 1983; Sequence of the hemagglutinin gene of influenza virus A/Memphis/1/71 and previously uncharacterised monoclonal antibody-derived variants. Virology 128:495–501
    [Google Scholar]
  16. Reichlin M. 1970; The distribution of specificity in rabbit antisera directed toward human immunoglobulins. Immunochemistry 7:15–27
    [Google Scholar]
  17. Skehel J. J., Bayley P. M., Brown E. B., Martin S. R., Waterfield M. D., White J. M., Wilson I. A., Wiley D. C. 1982; Changes in the conformation of influenza virus hemagglutinin at the pH optimum of virus-mediated membrane fusion. Proceedings of the National Academy of Sciences, U. S. A 79:968–972
    [Google Scholar]
  18. Ward C. W. 1981; Structure of the influenza virus hemagglutinin. Current Topics in Microbiology and Immunology 94/95:1–74
    [Google Scholar]
  19. Webster R. G., Brown L. E., Jackson D. C. 1983; Changes in the antigenicity of the hemagglutinin molecule of H3 influenza virus at acidic pH. Virology 126:587–599
    [Google Scholar]
  20. Wiley D. C., Skehel J. J., Waterfield M. D. 1977; Evidence from studies with a cross-linking reagent that the hemagglutinin of influenza virus is a trimer. Virology 79:446–448
    [Google Scholar]
  21. Wiley D. C., Wilson I. A., Skehel J. J. 1981; Structural identification of the antibody-binding sites of Hong Kong influenza haemagglutinin and their involvement in antigenic variation. Nature, London 289:373–378
    [Google Scholar]
  22. Wilson I. A., Skehel J. J., Wiley D. C. 1981; Structure of the haemagglutinin membrane glycoprotein of influenza virus at 3A resolution. Nature, London 289:366–373
    [Google Scholar]
  23. Yewdell J. W., Gerhard W., Bachi T. 1983; Monoclonal anti-hemagglutinin antibodies detect irreversible antigenic alterations that coincide with the acid activation of influenza virus A/PR/8/34-mediated hemolysis. Journal of Virology 48:239–248
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-66-8-1687
Loading
/content/journal/jgv/10.1099/0022-1317-66-8-1687
Loading

Data & Media loading...

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