1887

Abstract

SUMMARY

The possibility that the haemagglutinin of encephalomyocarditis () virus might be a glycoprotein was investigated by looking for sugars and amino sugars in purified EMC virus particles grown in Krebs ascitestumour cells in the presence of isotopically labelled glucosamine, acetyl glucosamine, galactosamine, mannosamine or fucose. With either isotopically labelled glucosamine or galactosamine the virus became significantly radioactive, whereas radioactivity was low in virusgrown in the presence of mannosamine, or fucose, possibly because the latter was poorly taken up by the cells; acetyl glucosamine was completely excluded by Krebs cells, so its ability to be incorporated into virus could not be tested.

Radioactivity derived from glucosamine remained firmly attached to the purified virus through both rate-zonal and equilibrium density centrifugation and chromatography on calcium phosphate. This radioactivity did not appear due to simple adsorption of either glucosamine or (after synthesis) acetyl glucosamine, or to contamination of the preparation with infected host cell membranes. Estimates of the number of glucosamine molecules incorporated into the virus had values which varied with the position of the isotope in the glucosamine added, suggesting that it was not incorporated unchanged.

Polyacrylamide gel electrophoresis of disrupted whole virus showed radio-activity derived from glucosamine to be present in all virus polypeptides, indicating that glucosamine was perhaps metabolized, amongst other things, to amino acids. Disruption of the virus with phenol revealed that 70 % of the radioactivity was in the virus , mostly in ribose, the remaining 30% being in virus protein, presumably as amino acids. The total amount of radioactivity that could be in the form of glucosamine did not allow for more than 0.6 glucosamine residues per virus particle. From this it was concluded that glucosamine is not a virus constituent that the virus particle does not contain glycoprotein and, consequently, that the virus haemagglutinin is not a glycoprotein.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-18-1-33
1973-01-01
2022-01-18
Loading full text...

Full text loading...

/deliver/fulltext/jgv/18/1/JV0180010033.html?itemId=/content/journal/jgv/10.1099/0022-1317-18-1-33&mimeType=html&fmt=ahah

References

  1. Bellett A. J. D., Burness A. T. H. 1963; Intracellular sites of synthesis of encephalomyocarditis virus components in Krebs-2 ascites tumour cells. Journal of General Microbiology 30:131–140
    [Google Scholar]
  2. Bruner R., Vinograd J. 1965; The evaluation of standard sedimentation coefficients of sodium RNA and sodium DNA from sedimentation velocity data in concentrated NaCl and CsCl solutions. Bio-chimica et biophysica acta 108:18–29
    [Google Scholar]
  3. Burness A. T. H. 1967; Separation of plaque-type variants of encephalomyocarditis virus by chromatography on calcium phosphate. Journal of Virology 1:308
    [Google Scholar]
  4. Burness A. T. H. 1969a; Purification and separation of encephalmoyocarditis virus variants by chromatography on calcium phosphate.. In Fundamental Techniques in Virology Edited by Habel K., Salzman N. New York and London: Academic Press;
    [Google Scholar]
  5. Burness A. T. H. 1969b; Purification of encephalomyocarditis virus. Journal of General Virology 5:291–303
    [Google Scholar]
  6. Burness A. T. H. 1970; Ribonucleic acid content of encephalomyocarditis virus. Journal of General Virology 6:373–380
    [Google Scholar]
  7. Burness A. T. H., Clothier F. W. 1970; Particle weight and other biophysical properties of encephalomyocarditis virus. Journal of General Virology 6:381–393
    [Google Scholar]
  8. Caliguiri L. A., Tamm I. 1970; The role of cytoplasmic membranes in poliovirus biosynthesis. Virology 42:100–111
    [Google Scholar]
  9. Compans R. W., Klenk H. D., Caliguiri 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]
  10. Davidson E. A. 1966; Analysis of sugars found in mucopolysaccharides. In Methods in Enzymology 8 Edited by Colowick S. P., Kaplan N. O., Menfeld E. F., Ginsberg V. New York and London: Academic Press;
    [Google Scholar]
  11. Dunker A. K., Rueckert R. R. 1969; Observations on molecular weight determinations on polyacrylamide gel. Journal of Biological Chemistry 244:5074–5080
    [Google Scholar]
  12. Fiske C. H., Subbarow Y. 1925; The colorimetric determination of phosphorous. Journal of Biological Chemistry 66:375
    [Google Scholar]
  13. Gierer A., Schramm G. 1956; Infectivity of ribonucleic acid from tobacco mosaic virus. Nature, London 177:702–703
    [Google Scholar]
  14. Goodheart C. R. 1965; A density mutant of encephalomyocarditis virus. Virology 26:466–477
    [Google Scholar]
  15. Gottschalk A. 1959; Chemistry of virus receptors. In The Viruses 351 Edited by Burnet F. M., Stanley W. M. New York and London: Academic Press;
    [Google Scholar]
  16. Halperen S., Stone H. O., Korant B. D. 1970 Isolation of glucosamine from the capsids of picornavirus. Abstract V18Bacteriological Proceedings, 70th Meeting of American Society for Microbiology Boston, Mass., U.S.A:
    [Google Scholar]
  17. Haslam E. A., Hampson A. W., Egan J. A., White D. O. 1970a; The polypeptides of influenza virus. II. Interpretation of polyacrylamide gel electrophoresis patterns. Virology 42:555–565
    [Google Scholar]
  18. Haslam E. A., hampson A. W., Radiskevis I., White D. O. 1970b; The polypeptides of influenza virus. III. Identification of the hemagglutinin, neuraminidase and nucleocapsid proteins. Virology 42:566–575
    [Google Scholar]
  19. Herberg R. J. 1960; Determination of carbon-14 and tritium in blood and other whole tissues. Analytical Chemistry 32:42–46
    [Google Scholar]
  20. Hoskins J. M., Sanders F. K. 1957; Propagation of mouse encephalomyocarditis virus in ascitestumour cells maintained in vitro. British Journal of Experimental Pathology 38:268–272
    [Google Scholar]
  21. International Critical Tables of Numerical Data, Physics, Chemistry, Technology; 1927; 2337 Edited by Washbum E. W. et al New York and London: McGraw-Hill;
    [Google Scholar]
  22. Klenk H. D., Caliguiri L. A., Choppin P. W. 1970; The proteins of the parainfluenza virus SV5. II. The carbohydrate content and glycoproteins of the virion. Virology 42:473–481
    [Google Scholar]
  23. Markham R., Smith J. D. 1952; The structure of ribonucleic acids. I. Cyclic nucleotides produced by ribonuclease and by alkaline hydrolysis. Biochemical Journal 52:552–557
    [Google Scholar]
  24. Peterson E. A., Sober H. A. 1959; Variable gradient device for chromatography. Analytical Chemistry 31:857–862
    [Google Scholar]
  25. Rueckert R. R. 1965; Studies on the structure of viruses of the Columbia SK Group. II. The protein sub-units of ME-virus and other members of the Columbia SK Group. Virology 26:345–358
    [Google Scholar]
  26. Rueckert R. R. 1971 Picornaviral architecture. In Comparative Virology Edited by Maramorosch K., Kurstak E. New York and London: Academic Press;
    [Google Scholar]
  27. Sanders F. K., Huppert J., Hoskins J. M. 1958; Replication of an animal virus. Symposium Society of Experimental Biology 12:123
    [Google Scholar]
  28. Schwerdt C. E. 1959 Quantitative relationships between virus particles and their functional activity.. In The Viruses 1329 Edited by Burnet F. M., Stanley W. M. New York and London: Academic Press;
    [Google Scholar]
  29. Strauss J. H., Burge B. W., Darnell J. E. 1969; Carbohydrate content of the membrane protein of Sindbis virus. Journal of Molecular Biology 47:437–448
    [Google Scholar]
  30. Tillotson J. R., Lerner A. M. 1966; Effect of periodate oxidation on haemagglutinating and antibodyproducing capacities of certain enteroviruses and reoviruses. Proceedings of the National Academy of Sciences of the United States of America 56:1143–1150
    [Google Scholar]
  31. Wheat R. W. 1966 Analysis of hexosamines in bacterial polysaccharides by chromatographic procedures. In Methods in Enzymology 8 Edited by Colowick S. P., Kaplan N. O., Menfeld E. F., Ginsberg V. New York and London: Academic Press;
    [Google Scholar]
  32. Wyatt G. R. 1955 Separation of nucleic acid components by chromatography on filter paper.. In The Nucleic Acids Edited by Chargaff E., Davidson J. N. New York and London: Academic Press;
    [Google Scholar]
  33. Young R. W., Fulhorst H. W. 1965; Recovery of S35radioactivity from protein-bearing polyacrylamide gel. Analytical Biochemistry 11:389
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-18-1-33
Loading
/content/journal/jgv/10.1099/0022-1317-18-1-33
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

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