1887

Abstract

SUMMARY

Protein IX from adenovirus type 2 was purified by two methods, one from groups of nine hexons obtained by disrupting purified virus by heating in the presence of deoxycholate, and the other by a previously published method. The purified protein was used to obtain a monospecific antiserum. Protein IX was found to possess both sub-group- and type-specific antigenic determinants which were apparently accessible within the groups of nine hexons. Approximately 15 molecules of IX were found per group of nine hexons and from considerations of symmetry it seemed possible that IX was located at the ‘corner to edge’ contacts between hexons in the icosahedron.

The protein in infected cells was found to possess approximately neutral charge as determined by immunoelectrophoresis. This was consistent with the amino acid composition, which showed it to be rich in serine, alanine and leucine with approximately half of its glutamic and aspartic acid residues amidified, and the isoelectric point of 6.0, as determined by two dimensional gel analysis. No free N-terminal amino acid was detectable. It is suggested that a unique tryptophan residue is located at around position 70 from the blocked N-terminus, on the basis of chemical cleavage by BNPS-skatole. Based on one tryptophan residue a total of 107 amino acids and a mol. wt. of 11200 was deduced. Analysis of S-methionine-labelled infected cell extracts in a two-dimensional gel system showed that the synthesis of polypeptide IX could be detected early in infection, i.e. in the presence of an inhibitor of DNA synthesis.

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1979-09-01
2022-08-13
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References

  1. Anderson C. W., Baum P. R., Gesteland R. F. 1973; Processing of adenovirus 2-induced proteins. Journal of Virology 12:241–252
    [Google Scholar]
  2. Blair G. E., Russell W. C. 1978; Identification of a protein kinase activity associated with human adenoviruses. Virology 86:157–166
    [Google Scholar]
  3. Bonner W. M., Laskey R. A. 1974; A film detection method for tritium labelled proteins and nucleic acids in polyacrylamide gels. European Journal of Biochemistry 46:83–88
    [Google Scholar]
  4. Boulanger P., Devaux C., Lemay P. 1978a; Isolation and characterization of a slow-migrating class of adenovirus type 2 hexons. Virology 84:456–468
    [Google Scholar]
  5. Boulanger P. A., Devaux C., Loucheux-Lefebvre M. H. 1978b; Comparative optical properties of free and assembled hexon capsomeres of human adenovirus type 2. FEBS Letters 85:52–56
    [Google Scholar]
  6. Boulanger P. A., Puvion F. 1973; Large-scale preparation of soluble adenovirus hexon, penton and fibre antigens in highly purified form. European Journal of Biochemistry 39:37–42
    [Google Scholar]
  7. Casola L., Di mattheo G., Di prisco G., Cervone F. 1974; Dansyl chloride binding to proteins. Quantitative estimation of N-terminal, lysyl and tyrosyl residues by the radioactive reagent. Analytical Biochemistry 57:38–45
    [Google Scholar]
  8. Crawford L. V., O’Farrell P. Z. 1978; The effect of alkylation on the physical properties of SV40 T-antigen species. Journal of Virology 29:587–596
    [Google Scholar]
  9. Crowther R. A., Franklin R. M. 1972; The structure of the groups of nine hexons from aaenovnua. Journal of Molecular Biology 68:181–184
    [Google Scholar]
  10. Everitt E., Philipson L. 1974; Structural proteins of adenoviruses. XI. Purification of three low molecular weight virion proteins of adenovirus type 2 and their synthesis during productive infection. Virology 62:253–269
    [Google Scholar]
  11. Everitt E., Sundquist B., Pettersson U., Philipson L. 1973; Structural proteins of adenoviruses. X. Isolation and topography of low molecular weight antigens from the virion of adenovirus type 2. Virology 52:130–147
    [Google Scholar]
  12. Everitt E., Lutter L., Philipson L. 1975; Structural proteins of adenoviruses. XII — Location and neighbour relationship among proteins of adenovirion type 2 as revealed by enzymatic iodination, immunoprecipitation and chemical cross-linking. Virology 67:197–208
    [Google Scholar]
  13. Eylar E. H., Sackson J. J., Bennet C. D., Kniskern P. J., Brostoff S. W. 1974; The chicken A1 protein. Phylogenetic variation in the amino acid sequence of the encephalitogenic site. Journal of Biological Chemistry 249:3710–3716
    [Google Scholar]
  14. Ginsberg H. S., Pereira H. G., Valentine R. C., Wilcox W. C. 1966; A proposed terminology for the adenovirus antigens and virion morphological subunits. Virology 28:782–783
    [Google Scholar]
  15. Green M. 1970; Oncogenic viruses. Annual Review of Biochemistry 39:701–756
    [Google Scholar]
  16. Green M., Pina M. 1963; Biochemical studies on the adenovirus multiplication. IV. Isolation, purification and chemical analysis of adenovirus. Virology 20:199–207
    [Google Scholar]
  17. Hunter W. M., Greenwood F. C. 1962; Preparation of iodine 125I-Iabelled human growth hormone of high specific activity. Nature, London 194:495–498
    [Google Scholar]
  18. Kessler S. 1975; Rapid isolation of antigens from cells with a Staphylococcal protein A antibody adsorbant: parameters of the interaction of antibody-antigen complexes with protein A. Journal of Immunology 115:1617–1624
    [Google Scholar]
  19. Laurell C. B. 1965; Antigen-antibody crossed immunoeleetrophoresis. Analytical Biochemistry to:358–361
    [Google Scholar]
  20. Lewis J. B., Anderson C. W., Atkins J. F., Gesteland R. F. 1975; The origin and destiny of adenovirus proteins. Cold Spring Harbor Symposium on Quantitative Biology 39:581–590
    [Google Scholar]
  21. Liu T. Y., Chang Y. H. 1971; Hydrolysis of proteins with p. toluene sulfonic acid. Determination of tryptophan. Journal of Biological Chemistry 246:2842–2848
    [Google Scholar]
  22. 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]
  23. Maizel J. V., White D. O., Sharff M. D. 1968; The polypeptides of adenovirus. II. Soluble proteins, cores, top components and the structure of the virion. Virology 36:126–136
    [Google Scholar]
  24. Martin G. R., Warocquier R., Boulanger P. A. 1975; Quantitation of adenovirus soluble antigens by crossed immunoelectrophoresis: application to serological characterization of mutants. Intervirology 5:62–172
    [Google Scholar]
  25. Nermut M. V. 1975; Fine structure of adenovirus type 5-I. Virus capsid. Virology 65:480–495
    [Google Scholar]
  26. O’Farrell P. H. 1975; High resolution two-dimensional electrophoresis of proteins. Journal of Biological Chemistry 250:4007–4021
    [Google Scholar]
  27. Pereira H. G., Wrigley N. G. 1974; In vitro reconstitution, hexon bonding and handedness of incomplete adenovirus. Journal of Molecular Biology 85:617–630
    [Google Scholar]
  28. Persson H., Pettersson U., Mathews M. B. 1978; Synthesis of a structural adenovirus polypeptide in the absence of viral DNA replication. Virology 90:67–79
    [Google Scholar]
  29. Petterson U., Mathews M. B. 1977; The gene and messenger RNA for adenovirus polypeptide IX. Cell 12:741–750
    [Google Scholar]
  30. Philipson L., Lindberg U. 1974; Reproduction of adenoviruses. In Comprehensive Virology vol 3 pp 143–227 Edited by Fraenkel-Conrat H., Wagner R. R. New York: Plenum Press;
    [Google Scholar]
  31. Prage L., Pettersson U., Hoglund S., Lonberg-Holm K., Philipson L. 1970; Structural proteins of adenoviruses. IV. Sequential degradation of the adenovirus type 2 virion. Virology 42:341–358
    [Google Scholar]
  32. Russell W. C., Blair G. E. 1977; Polypeptide phosphorylation in adenovirus-infected cells. Journal of General Virology 34:19–35
    [Google Scholar]
  33. Russell W. C., McIntosh K., Skehel J. J. 1971; The preparation and properties of adenovirus cores. Journal of General Virology 11:35–46
    [Google Scholar]
  34. Russell W. C., Skehel J. J. 1972; The polypeptides of adenovirus infected cells. Journal of General Virology 15:45–57
    [Google Scholar]
  35. Smith K. O., Gehle W. D., Trousdale M. D. 1965; Architecture of the adenovirus capsid. Journal of Bacteriology 90:254–261
    [Google Scholar]
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