1887

Abstract

SUMMARY

An assay system has been developed for the adenovirus endoproteinase which utilizes the synthetic peptide MSGGAFSW, derived from the cleavage site of the adenovirus type 2 (Ad2) protein pVI. MSGGAFSW was shown to be cleaved at the G–A bond when incubated with a source of Ad2 proteinase. Digestions were readily monitored by either fast protein liquid chromatography or thin layer electrophoresis, enabling the rapid production of quantitative data. Comparison of the peptide assay with a previously described [S]methionine assay system showed it to be faster, cleaner and less prone to extreme conditions of pH and ionic strength. The effect on adenovirus proteinase activity of a number of inhibitors was assessed using both the [S]methionine and peptide assays. Identical inhibitor profiles were obtained and these suggested that the adenovirus enzyme is a cysteine proteinase. The 23K gene product, thought to be the proteinase, contains cysteine and histidine residues at positions 122 and 54, respectively, that could constitute part of the active site of a cysteine proteinase. These amino acids and their surrounding residues are conserved in all serotypes examined and appear to bear some resemblance to those in the putative active sites of the 3C proteinases in picornaviruses.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-70-12-3215
1989-12-01
2024-12-03
Loading full text...

Full text loading...

/deliver/fulltext/jgv/70/12/JV0700123215.html?itemId=/content/journal/jgv/10.1099/0022-1317-70-12-3215&mimeType=html&fmt=ahah

References

  1. Argos P., Kamer G., Nicklin M. J. H., Wimmer E. 1984; Similarity in gene organisation and homology between proteins of animal picornaviruses and a plant comovirus suggest common ancestry of these virus families. Nucleic Acids Research 12:7251–7267
    [Google Scholar]
  2. Atherton E., Cameron L. R., Sheppard R. C. 1988; Peptide synthesis. Part 10. Use of pentafluorophenyl esters of fluorenyl methoxycarbonylamino acids in solid phase synthesis. Tetrahedron 44:843–857
    [Google Scholar]
  3. Barrett A. J. 1977; Introduction to the history and classification of tissue proteinases. In Proteinases in Mammalian Cells and Tissues1–55 Barrett A. J. Amsterdam: North-Holland:
    [Google Scholar]
  4. Bazan J. F., Fletterick R. J. 1988; Viral cysteine proteases are homologous to the trypsin-like family of serine proteases: structural and functional implications. Proceedings of the National Academy of SciencesU.S.A 85:7872–7876
    [Google Scholar]
  5. Bazan J. F., Fletterick R. J. 1989; Detection of a trypsin-like serine protease domain in flaviviruses and pestiviruses. Virology 171:637–639
    [Google Scholar]
  6. Bhatti A. R., Weber J. 1979a; Protease of adenovirus type 2: subcellular localization. Journal of Biological Chemistry 254:12265–12268
    [Google Scholar]
  7. Bhatti A. R., Weber J. 1979b; Protease of adenovirus type 2: partial characterization. Virology 96:478–485
    [Google Scholar]
  8. Bond J. S., Butler P. E. 1987; Intracellular proteases. Annual Review of Biochemistry 56:333–364
    [Google Scholar]
  9. Brenner S. 1988; The molecular evolution of genes and proteins: a tale of two serines. Nature, London 334:528–530
    [Google Scholar]
  10. Coulson A. F. W., Collins J. F., Lyall A. 1987; Protein and nucleic acid sequence database searching: a suitable case for parallel processing. Computer Journal 30:420–424
    [Google Scholar]
  11. Geiger R., Fritz H. 1984; Trypsin. In Bergmeyer Methods of Enzymatic Analysis 5119–129 Fritz H. Weinheim: Verlag Chemie;
    [Google Scholar]
  12. Gorbalenya A. E., Donchenko A. P., Blinov V. M., Koonin E. V. 1989; Cysteine proteases of positive strand RNA viruses and chymotrypsin-like serine proteases. FEBS Letters 243:103–114
    [Google Scholar]
  13. Houde A., Weber J. M. 1987; Sequence of the protease of human subgroup E adenovirus type 4. Gene 54:51–56
    [Google Scholar]
  14. Jacobson M. F., Asso J., Baltimore D. 1970; Further evidence on the formation of poliovirus proteins. Journal of Molecular Biology 49:657–669
    [Google Scholar]
  15. Kotler M., Katz R. A., Danho W., Leis J., Skalka A. M. 1988; Synthetic peptides as substrates and inhibitors of a retroviral protease. Proceedings of the National Academy of SciencesU.S.A 85:4185–4189
    [Google Scholar]
  16. Krausslich H., Wimmer E. 1988; Viral proteinases. Annual Review of Biochemistry 57:701–754
    [Google Scholar]
  17. Nicklin M. J. H., Toyoda H., Murray M. G., Wimmer E. 1986; Proteolytic processing of polio and related viruses. Bio/Technology 4:36–42
    [Google Scholar]
  18. Roth M. 1971; Fluorescence reaction of amino acids. Analytical Chemistry 43:880–882
    [Google Scholar]
  19. Russell W. C., Blair G. E. 1977; Polypeptide phosphorylation in adenovirus-infected cells. Journal of General Virology 34:19–35
    [Google Scholar]
  20. Russell W. C., Valentine R. C., Pereira H. G. 1967; The effect of heat on the anatomy of the adenovirus. Journal of General Virology 1:509–522
    [Google Scholar]
  21. Toyoda H., Nicklin M. J. H., Murray M. G., Anderson C. W., Dunn J. J., Studier F. W., Wimmer E. 1986; A second virus-encoded proteinase involved in proteolytic processing of poliovirus polyprotein. Cell 45:761–770
    [Google Scholar]
  22. Tremblay M. L., Dery C. V., Talbot B. G., Weber J. 1983; In vitro cleavage specificity of the adenovirus type 2 proteinase. Biochimica et biophysica acta 743:239–245
    [Google Scholar]
  23. Vos H. L., Van Der Lee F. M., Reemst A. M. C. B., Van Loon A. E., Sussenbach J. S. 1988; The genes encoding the DNA binding protein and the 23K protease of adenovirus types 40 and 41. Virology 163:1–10
    [Google Scholar]
  24. Weber J. 1976; Genetic analysis of adenovirus type 2. III. Temperature sensitivity of processing of viral proteins. Journal of Virology 17:462–471
    [Google Scholar]
  25. Weber J. M., Houde A. 1987; Spontaneous reversion of a C/T transition mutation in the adenovirus endoproteinase gene. Virology 156:427–428
    [Google Scholar]
  26. Webster A., Russell S., Talbot P., Russell W. C., Kemp G. D. 1989; Characterization of the adenovirus proteinase: substrate specificity. Journal of General Virology 70:3225–3234
    [Google Scholar]
  27. Whittaker J. R., Perez-Villasenor J. 1968; Chemical modification of papain. 1. Reaction with the chloromethyl ketones of phenylalanine and lysine and with phenylmethylsulphonyl fluoride. Archives of Biochemistry and Biophysics 124:70–78
    [Google Scholar]
  28. Willenbrock F., Brocklehurst K. 1985; A general framework of cysteine proteinase mechanism deduced from studies on enzymes with structurally different analogous catalytic-site residues Asp-158 and -161 (papain and actinidin), Gly-196 (cathepsin B) and Asn-165 (cathepsin H). Biochemical Journal 227:521–528
    [Google Scholar]
  29. Yeh-Kai L., Akusjarvi G., Alestrom P., Pettersson U., Tremblay M., Weber J. M. 1983; Genetic identification of an endoproteinase encoded by the adenovirus genome. Journal of Molecular Biology 167:217–222
    [Google Scholar]
/content/journal/jgv/10.1099/0022-1317-70-12-3215
Loading
/content/journal/jgv/10.1099/0022-1317-70-12-3215
Loading

Data & Media loading...

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