1887

Abstract

SUMMARY

Haemagglutinating encephalomyelitis virus (HEV), a member of the coronavirus family, was purified and analysed by SDS-polyacrylamide gel electrophoresis. It was shown to contain eight polypeptides, seven of which were glycosylated. They had apparent mol. wt. of 180000 (GP 180), 130000 (GP 130), 120000 (GP 120) 76000 (GP 76), 64000 (VP 64), 54000 (GP 54), 32000 (GP 32) and 31000 (GP 31). Electrophoresis of virus samples dissociated under varying conditions showed that GP 54 and GP 120 could be interpreted as larger products of GP 31 and GP 32 and of GP 76, respectively. GP 76 also appeared as a dimer with a mol. wt. of 140000 (GP 140) in the absence of β-mercaptoethanol. Subviral particles, obtained by treatment with bromelain, banded at a slightly lower density than the intact virus and lacked surface projections. Analysis of these particles indicated that GP 180, GP 130 and GP 76 are associated with the virus projections. A small part of GP 31 and GP 32 also appeared to protrude from the lipid envelope, since 20% of each molecule was sensitive to digestion. Two glycoproteins, GP 130 and GP 76, were solubilized with the detergent Triton X-100 and separated by rate zonal centrifugation. According to its activity in indirect haemagglutination tests, GP 76 was considered to be a monovalent haemagglutinin subunit.

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1980-05-01
2024-03-29
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References

  1. Andrif S. K., Pensaert M., Callebaut P. 1978; Pathogenicity of hemagglutinating encephalomyelitis (vomiting and wasting disease) virus of pigs, using different routes of inoculation. Zentralblatt fiir Veterindrmedizin B 25:461–468
    [Google Scholar]
  2. Bernardi O. 1971; Chromatography of proteins on hydroxyapatite. In Methods in Enzymology vol 22 pp 325–339 Edited by Colowick S. P., Kaplan N. O. New York: Academic Press;
    [Google Scholar]
  3. Bingham R. W. 1975; The polypeptide composition of avian infectious bronchitis virus. Archives of Virology 49:207–216
    [Google Scholar]
  4. Compans R. W., Klenk H. D., Caltguirr L. A., Choppin P. W. 1970; Influenza virus proteins. I. Analysis of polypeptides of the virion and identification of spike glycoproteins. Virology 42:880–889
    [Google Scholar]
  5. Collins M. S., Alexander D. J., Harkness J. W. 1976; Heterogeneity of infectious bronchitis virus grown in eggs. Archives of Virology 50:55–72
    [Google Scholar]
  6. Garwes D. J., Pocock D. H. 1975; The polypeptide structure of transmissible gastroenteritis virus. Journal of General Virology 29:25–34
    [Google Scholar]
  7. Gschwender H. H., Rutter C., Popescu M. 1975; Use of iodinated organic compounds for the density gradient centrifugation of viruses. Archives of Virology 49:359–364
    [Google Scholar]
  8. Helenius A., Simons K. 1972; The binding of detergents to lipophilic and hydrophilic proteins. Journal of Biological Chemistry 247:3656–3661
    [Google Scholar]
  9. Hierholzer J. C. 1976; Purification and biophysical properties of human coronavirus 229E. Virology 75:155–165
    [Google Scholar]
  10. Hierholzer J. C., Palmer E. L., Whitfield S. G., Kaye H. S., Dowdle W. R. 1972; Protein composition of coronavirus OC43. Virology 48:516–527
    [Google Scholar]
  11. Lowry O. H., Rosebrough N. J., Farr A. L., Randall R. J. 1951; Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193:265–275
    [Google Scholar]
  12. Macnaughton M. R., Madge M. H. 1977; The polypeptide composition of avian infectious bronchitis virus particles. Archives of Virology 55:47–54
    [Google Scholar]
  13. Maizel J. V. Jun 1971; Polyacrylamide gel electrophoresis of viral proteins. In Methods in Virology vol 5 pp 179–246 Edited by Maramorosch K., Koprowski K. New York: Academic Press;
    [Google Scholar]
  14. Pensaert M. B., Callebaut P. E. 1974; Characteristics of a coronavirus causing vomiting and wasting in pigs. Archiv für die gesamte Virusforschung 44:35–50
    [Google Scholar]
  15. Pocock D. H. 1978; Effects of sulfhydryl reagents on the biological activities, polypeptide composition and morphology of haemagglutinating encephalomyelitis virus. Journal of General Virology 40:93–101
    [Google Scholar]
  16. Pocock D. H., Garwes D. J. 1977; The polypeptides of haemagglutinating encephalomyelitis virus and isolated subviral particles. Journal of General Virology 37:487–499
    [Google Scholar]
  17. Scheid A., Caliguiri L. A., Compans R. W., Choppin P. W. 1972; Isolation of paramyxovirus glycoproteins. Association of both hemagglutinating and neuraminidase activities with the larger SV 5 glycoprotein. Virology 50:640–652
    [Google Scholar]
  18. Shapiro A. L. 1967; Molecular weight estimation of polypeptide chains by electrophoresis in SDS-polyacrylamide gels. Biochemical and Biophysical Research Communications 28:814–820
    [Google Scholar]
  19. Sturman L. S. 1977; Characterization of a coronavirus. I. Structural proteins: effect of preparative conditions on the migration of protein in polyacrylamide gels. Virology 77:637–649
    [Google Scholar]
  20. Tyrrell D. A., Almeida J. D., Cunningham C. H., Dowdle W. R., Hofstad M. S., Macintosh K., Tajima M., Zakstelskaya L. Ya., Easterday B. C., Kapikian A., Bingham R. W. 1975; Coronaviridae. Intervirology 5:76–82
    [Google Scholar]
  21. Uriel J., Avrameas S., Grabar P. 1964; Caractérisation d’enzymes après électrophorèse et analyse immuno-électrophorétique en agarose. In Protides of the Biological Fluids XI Colloquium, Bruges 1963 pp 355–359 Edited by Peeters H. Amsterdam: Elsevier;
    [Google Scholar]
  22. Weber K., Osborn M. 1969; The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. Journal of Biological Chemistry 244:4406–4412
    [Google Scholar]
  23. Wege H., Wege H., Nagashima K., Ter Meulen V. 1979; Structural polypeptides of the murine coronavirus JHM. Journal of General Virology 42:37–47
    [Google Scholar]
  24. Zacharius R. M., Zell T. E., Morrison J. H., Woodlock J. J. 1969; Glycoprotein staining following electrophoresis on acrylamide gels. Analytical Biochemistry 30:148–152
    [Google Scholar]
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