1887

Abstract

The hepatitis B virus particle consists of an envelope carrying the surface antigen of the virus and an internal capsid consisting of the core antigen (HBcAg). The internal capsid contains the circular, partially dsDNA genome and the viral polymerase. Empty core particles have been produced in cells using a recombinant baculovirus vector, YMIKTc, that expresses a 21· 4K derivative of the HBcAg gene. The particles have been purified to homogeneity by caesium chloride density gradient centrifugation followed by glycerol gradient centrifugation. Physicochemical analysis of the core particles showed that they exhibited a sedimentation coefficient (s ,) of 82·5S and a diffusion coefficient () of 1 ·28 × 10 cm/s. The obtained by substitution of these values in the Svedberg equation was 5·8 × 10, using a partial specific volume of 0·73 ml/g for the viral protein as estimated from the amino acid composition. The determined from sedimentation equilibrium analyses was 6·3 × 10. Spectrophotometric and metabolic labelling analyses failed to detect nucleic acids in the core preparations. The data are at variance with the prediction that cores exhibit a T=3 symmetry and contain some 180 subunits. The results suggest that the baculovirus- expressed cores may contain up to 300 subunits of HBcAg protein.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-71-11-2755
1990-11-01
2021-10-25
Loading full text...

Full text loading...

/deliver/fulltext/jgv/71/11/JV0710112755.html?itemId=/content/journal/jgv/10.1099/0022-1317-71-11-2755&mimeType=html&fmt=ahah

References

  1. Albin C., Robinson W. S. 1980; Protein kinase activity in hepatitis B virus. Journal of Virology 34:297–302
    [Google Scholar]
  2. Argos P., Fuller S. 1988; A model for the hepatitis B virus core: prediction of antigenic sites and relationship to RNA virus capsid proteins. EMBO Journal 7:819–824
    [Google Scholar]
  3. Bowen T. J. 1970 An Introduction to Ultracentrifugation London: John Wiley;
    [Google Scholar]
  4. Cohn E. J., Edsall J. T. 1943 Proteins, Amino Acids and Peptides pp. 275–377 American Chemical Society Monograph Series New York: Hafner Publishing Co;
    [Google Scholar]
  5. Dane D. S., Cameron C. H., Briggs A. 1970; Virus-like particles in serum of patients with Australian antigen-associated hepatitis. Lancet i:695–698
    [Google Scholar]
  6. Edelstein S. J., Schachman H. K. 1973; Measurement of partial specific volume by sedimentation equilbrium in H2O-D2O solutions. Methods in Enzymology 27D:82–98
    [Google Scholar]
  7. Gagan W. L. 1966; The significance of the “partial specific volume” obtained from sedimentation data. Biochemistry 5:2553–2557
    [Google Scholar]
  8. Kaplan P. N., Greenman R. L., Gerin I. L., Purcell R. H., Robinson W. L. 1973; DNA polymerase associated with human hepatitis B antigen. Journal of Virology 12:995–1005
    [Google Scholar]
  9. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature; London: 227680–685
    [Google Scholar]
  10. Ono Y., Onda H., Sasada R., Igarashi K., Sugino Y., Nishioka K. 1983; The complete nucleotide sequences of the cloned hepatitis B virus DNA; subtypes adr and adw . Nucleic Acids Research 11:1747–1757
    [Google Scholar]
  11. Pasek M., Goto T., Gilbert W., Zink B., Schaller H., Mackay P., Leadbetter G., Murray K. 1979; Hepatitis B virus genes and their expression in E. coli . Nature; London: 282575–579
    [Google Scholar]
  12. Takehara K., Ireland D., Bishop D. H. L. 1988; Co-expression of the hepatitis B surface and core antigens using baculovirus multiple expression vectors. Journal of General Virology 69:2763–2777
    [Google Scholar]
  13. Tiollais P., Purcell C., Dejean A. 1985; The hepatitis B virus. Nature; London: 317489–495
    [Google Scholar]
  14. Warburg O., Christian W. 1941; Isolierung und Kristallisation des Gärungsferments Enolase. Biochemische Zeitschrift 310:384–421
    [Google Scholar]
  15. Yphantis D. A. 1964; Equilibrium ultracentrifugation of dilute solutions. Biochemistry 3:297–317
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-71-11-2755
Loading
/content/journal/jgv/10.1099/0022-1317-71-11-2755
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