1887

Abstract

Summary

Proteolytic digestion of purified whole tick-borne encephalitis virus or its isolated envelope glycoprotein (E) in the form of rosettes yields an 9000 fragment that is resistant to further digestion and carries polyclonal and monoclonal antibody-defined antigenic determinants. In a denaturation/renaturation experiment it was demonstrated that the antigenic reactivity of this domain, which was lost upon reduction and carboxymethylation, could be regained if the reducing agent was dialysed out before carboxymethylation. By the use of [S]cysteine-labelled E protein and amino acid analysis it was confirmed that the reacquisition of antigenic reactivity in the renaturation experiment was associated with the reformation of disulphide bridges, which apparently confer structural stability to this part of the molecule. By the experiments performed we have identified an independently folding antigenically active domain of the E protein that is stabilized by disulphide bridges and has a strong tendency for renaturation.

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1987-08-01
2024-12-10
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References

  1. Dalgarno L., Trent D. W., Strauss J. H., Rice C. M. 1986; Partial nucleotide sequence of the Murray Valley encephalitis virus genome. Comparison of the encoded polypeptides with yellow fever virus structural and non-structural proteins. Journal of Molecular Biology 187:309–323
    [Google Scholar]
  2. Della-Porta A. J., Westaway E. G. 1911; Immune response in rabbits to virion and nonvirion antigens of the flavivirus Kunjin. Infection and Immunity 15:874–882
    [Google Scholar]
  3. Hawkes R. 1982; Identification of concanavalin A-binding proteins after sodium dodecyl sulfate-gel electrophoresis and protein blotting. Analytical Biochemistry 123:143–146
    [Google Scholar]
  4. Heinz F. X., Kunz C. 1980; Formation of polymeric glycoprotein complexes from a flavivirus: tick-borne encephalitis. Journal of General Virology 49:125–132
    [Google Scholar]
  5. Heinz F. X., Kunz C. 1981; Homogeneity of the structural glycoprotein from European isolates of tick-borne encephalitis virus: comparison with other flaviviruses. Journal of General Virology 57:263–274
    [Google Scholar]
  6. Heinz F. X., Tuma W., Kunz C. 1981; Antigenic and immunogenic properties of defined physical forms of tickborne encephalitis virus structural proteins. Infection and Immunity 33:250–257
    [Google Scholar]
  7. Heinz F. X., Berger R., Tuma W., Kunz C. 1983a; . A topological and functional model of epitopes on the structural glycoprotein of tick-bome encephalitis virus defined by monoclonal antibodies Virology 126:525–537
    [Google Scholar]
  8. Heinz F. X., Berger R., Tuma W. 1983b; Location of immunodominant antigenic determinants on fragments of the tick-borne encephalitis virus glycoprotein. Evidence for two different mechanisms by which antibodies mediate neutralization hemagglutination inhibition. Virology 130:485–501
    [Google Scholar]
  9. Heinz F. X., Tuma W., Guirakhoo F., Berger R., Kunz C. 1984; Immunogenicity of tick-borne encephalitis virus glycoprotein fragments: epitope-specific analysis of the antibody response. Virology 65:1921–1929
    [Google Scholar]
  10. Kitano T., Suzuki K., Yamoguchi T. 1974; Morphological chemical and biological characterization of Japanese encephalitis virus virion and its hemagglutinin. Journal of Virology 14:631–639
    [Google Scholar]
  11. Laemmli U. K., Favre M. 1973; Maturation of the head of bacteriophage T4.1. DNA packaging events. Journal of Molecular Biology 80:575–599
    [Google Scholar]
  12. Nowak T., Wengler G. 1987; Analysis of disulfides present in the membrane proteins of the West Nile flavivirus. Virology 156:127–137
    [Google Scholar]
  13. Pletnev A. G., Yamshchikov V. F., Blinov V. M. 1986; Tick-borne encephalitis virus genome. The nucleotide sequence coding for virion structural proteins. FEBS Letters 200:317–321
    [Google Scholar]
  14. Rice C. M., Lenches E. M., Eddy S. R., Shin S. J., Sheets R. L., Strauss J. H. 1985; Nucleotide sequence of yellow fever virus: implications for flavivirus gene expression and evolution. Science 229:726–733
    [Google Scholar]
  15. Shiroishi T., Evans G. A., Appella E., Ozato K. 1984; Role of a disulfide bridge in the immune function of major histocompatibility class I antigen as studied by in vitro mutagenesis. Proceedings of the National Academy of Sciences U.S.A.: 817544–7548
    [Google Scholar]
  16. Towbin H., Staehelin T., Gordon J. 1979; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences U.S.A.: 764350–4354
    [Google Scholar]
  17. Trent D. W. 1977; Antigenic characterization of flavivirus structural proteins separated by isoelectric focusing. Journal of Virology 22:608–618
    [Google Scholar]
  18. Webster R. G., Laver W. G., Air G. M., Schild G. C. 1982; Molecular mechanisms of variation in influenza viruses. Nature; London: 296115–121
    [Google Scholar]
  19. Wengler G., Castle E., Leidner U., Nowak T., Wengler G. 1985; Sequence analysis of the membrane protein V3 of the flavivirus West Nile virus and of its gene. Virology 147:264–274
    [Google Scholar]
  20. Westaway E. G., Brinton M. A., Gaidamovich S. Y., Horzinek M. C., Igarashi A., Kääriäinen L., Lvov D. K., Porterfield J. S., Russell P. K., Trent D. W. 1985; Flaviviridae. Intervirology 24:183–192
    [Google Scholar]
  21. Winkler G., Heinz F. X., Guirakhoo F., Kunz C. 1985; Separation of flavivirus membrane and capsid proteins by multistep high performance liquid chromatography optimized by immunological monitoring. Journal of Chromatography 326:113–119
    [Google Scholar]
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