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Volume 65, Issue 2

Purification, Morphology and Antigenic Characterization of Measles Virus Envelope Components Free

Abstract

SUMMARY

Measles virus haemagglutinin (H), fusion (F) and matrix (M) components were purified by affinity chromatography using monoclonal antibodies coupled to CNBr-activated Sepharose. H and M proteins were purified to homogeneity as determined by polyacrylamide gel electrophoresis by a single cycle of adsorption-desorption. The corresponding purification of the F protein required two cycles of adsorption-desorption. After the second adsorption an extra wash with 1 -guanidine-HCl was employed to remove the contaminating cellular actin. Electron microscopic examination of the purified envelope proteins showed that, at pH 6.0, the H peplomers had a truncated conical shape (width 6.5 to 4 nm, length 16 nm), and the F peplomers had a club-like shape (dimensions of the oval head 6 × 9 nm, length 15 nm). Lengths of both peplomers were measured excluding their undiscernible hydrophobic part. The M component at pH 3.0 appeared as a rounded particle (diam. 8 nm, central accumulation of contrast 1.5 nm) suggested to include four to six M polypeptides. Rabbit hyperimmune sera were prepared against all three purified envelope components. These sera reacted only with the homologous antigen in radioimmunoprecipitation assays. Both antisera against the H and F components neutralized the virus and blocked virus-specific haemolysis, but only anti-H serum inhibited haemagglutination.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): envelope proteins , isolation , measles virus and morphology and antigenicity
© Journal of General Virology 1984
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1984-02-01
2024-04-19
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References

  1. Bellini W. J., McFarlin D. E., Silver G. D., Mingioli E. S., McFarland H. F. 1981; Immune reactivity of the purified hemagglutinin of measles virus. Infection and Immunity 32:1051–1057
     [Google Scholar]
  2. Birrer M. J., Udem S., Nathenson S., Bloom B. R. 1981; Antigenic variants of measles virus. Nature, London 293:67–69
     [Google Scholar]
  3. Bonner W. M., Laskey R. A. 1974; A film detection method for tritium-labeled proteins and nucleic acids in polyacrylamide gels. European Journal of Biochemistry 46:83–88
     [Google Scholar]
  4. Casali P., Sissons J. G. P., Fujinami R. S., Oldstone M. B. A. 1981; Purification of measles virus glycoproteins and their integration into artificial lipid membranes. Journal of General Virology 54:161–171
     [Google Scholar]
  5. Christie M., Endresen C., Haukenes G. 1981; Purification of measles virus H polypeptide and of F polypeptide. Archives of Virology 69:177–187
     [Google Scholar]
  6. Fujinami R. S., Sissons J. G. P., Oldstone M. B. A. 1981; Immunereactive measles virus polypeptides on the cell’s surface: turnover and relationship of the glycoproteins to each other and to HLA determinants. Journal of Immunology 127:935–940
     [Google Scholar]
  7. Giuffre R. M., Tovell D. R., Kay C. M., Tyrrell D. L. 1982; Evidence for an interaction between the membrane protein of a paramyxovirus and actin. Journal of Virology 42:963–968
     [Google Scholar]
  8. Graves M. C. 1981; Measles virus polypeptides in infected cells studied by immune precipitation and one-dimensional peptide mapping. Journal of Virology 38:224–230
     [Google Scholar]
  9. Hall W. W., Kiessling W., Ter Meulen V. 1978; Membrane proteins of subacute sclerosing panencephalitis and measles viruses. Nature, London 272:460–462
     [Google Scholar]
  10. Hardwick J. M., Bussell R. H. 1978; Glycoproteins of measles virus under reducing and nonreducing conditions. Journal of Virology 25:687–692
     [Google Scholar]
  11. Heggeness M. H., Smith P. R., Choppin P. W. 1982; In vitro assembly of the nonglycosylated membrane protein (M) of Sendai virus. Proceedings of the National Academy of Sciences, U. S. A. 79:6232–6236
     [Google Scholar]
  12. Hewitt J. A., Nermut M. V. 1977; A morphological study of the M protein of Sendai virus. Journal of General Virology 34:127–136
     [Google Scholar]
  13. Iinuma M., Yoshida T., Nagai Y., Maeno K., Matsumoto T. 1971; Subunits of NDV. Hemagglutinin and neuraminidase subunits of Newcastle disease virus. Virology 46:663–677
     [Google Scholar]
  14. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
     [Google Scholar]
  15. Lazarides E., Lindberg U. 1974; Actin is the naturally occurring inhibitor of deoxyribonuclease I. Proceedings of the National Academy of Sciences, U. S. A. 71:4742–4746
     [Google Scholar]
  16. Lund G. A., Salmi A. A. 1981; Purification and characterization of measles virus haemagglutinin protein G. Journal of General Virology 56:185–193
     [Google Scholar]
  17. Merz D. C., Scheid A., Choppin P. W. 1980; Importance of antibodies to the fusion glycoproteins of paramyxoviruses in the prevention of spread of infection. Journal of Experimental Medicine 151:275–288
     [Google Scholar]
  18. Merz D. C., Scheid A., Choppin P. W. 1981; Immunological studies of the functions of paramyxovirus glycoproteins. Virology 109:94–105
     [Google Scholar]
  19. Mountcastle W. E., Choppin P. W. 1977; A comparison of the polypeptides of four measles virus strains. Virology 78:463–474
     [Google Scholar]
  20. Norrby E., Appel M. J. 1980; Humoral immunity to canine distemper after immunization of dogs with inactivated and live measles virus. Archives of Virology 66:169–177
     [Google Scholar]
  21. Norrby E., Gollmar Y. 1972; Appearance and persistence of antibodies against different virus components after regular measles infection. Infection and Immunity 6:240–247
     [Google Scholar]
  22. Norrby E., Gollmar Y. 1975; Identification of measles virus-specific hemolysis-inhibiting antibodies separate from hemagglutinating-inhibiting antibodies. Infection and Immunity 11:231–239
     [Google Scholar]
  23. Norrby E., Enders-Ruckle G., Ter Meulen V. 1975; Differences in the appearance of antibodies to structural components of measles virus after immunization with inactivated and live virus. Journal of Infectious Diseases 132:262–269
     [Google Scholar]
  24. Norrby E., Chen S.-N., Togashi T., Sheshberadaran H., Johnson K. P. 1982; Five measles virus antigens demonstrated by use of mouse hybridoma antibodies in productively infected tissue culture cells. Archives of Virology 71:1–11
     [Google Scholar]
  25. Örvell C., Norrby E. 1980; Immunological relationships between homologous structural polypeptides of measles and canine distemper virus. Journal of General Virology 50:231–245
     [Google Scholar]
  26. Scheid A., Caliguiri L. A., Comp Ans R. W., Choppin P. W. 1972; Isolation of paramyxovirus glycoproteins. Association of both hemagglutinating and neuraminidase activities with the larger SV5 glycoprotein. Virology 50:640–652
     [Google Scholar]
  27. Sheshberadaran H., Chen S-N., Norrby E. 1983; Monoclonal antibodies against five structural components of measles virus. I. Characterization of antigenic determinants on nine strains of measles virus. Virology 128:341–353
     [Google Scholar]
  28. Shimizu K., Shimizu Y. K., Kohama T., Ishida N. 1974; Isolation and characterization of two distinct types of HVJ (Sendai virus) spikes. Virology 62:90–101
     [Google Scholar]
  29. Togashi T., Orvell C., Vartdal F., Norrby E. 1981; Production of antibodies against measles virions by use of the mouse hybridoma technique. Archives of Virology 67:149–157
     [Google Scholar]
  30. Trudgett A., Gould E. A., Armstrong M., Mingioli E. S., McFarlin D. E. 1981; Antigenic differences in the hemagglutinin of measles and related viruses. Virology 109:180–182
     [Google Scholar]
  31. Tyrrell D. L. J., Norrby E. 1978; Structural polypeptides of measles virus. Journal of General Virology 39:219–229
     [Google Scholar]
  32. Tyrrell D. L. J., Rafter D. J., Orvell C., Norrby E. 1980; Isolation and immunological characterization of the nucleocapsid and membrane proteins of measles virus. Journal of General Virology 51:307–315
     [Google Scholar]
  33. Wechsler S. L., Meissner H. C. 1982; Measles and SSPE viruses: similarities and differences. Progress in Medical Virology 28:65–95
     [Google Scholar]
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Purification, Morphology and Antigenic Characterization of Measles Virus Envelope Components
J. Gen. Virol. 65, 355 (1984); https://doi.org/10.1099/0022-1317-65-2-355
/content/journal/jgv/10.1099/0022-1317-65-2-355
/content/journal/jgv/10.1099/0022-1317-65-2-355
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