Skip to main content
SpringerLink
Log in

Studies on benzhydrazone, a specific inhibitor of herpesvirus glycoprotein synthesis. Size distribution of glycopeptides and endo-β-N-acetylglucosaminidase-H treatment

  • Original Papers
  • Published:
Archives of Virology Aims and scope Submit manuscript

Summary

Benzhydrazone is a bis-amidinohydrazone derivative which specifically hinders the formation of herpes simplex virus (HSV) glycoproteins. In this study we present some structural features of the oligosaccharide chains of herpesvirus glycoproteins synthesized in cells incubated with the inhibitor. Gel filtration analysis of glycopeptides, obtained through exhaustive pronase-digestion of infected cells after a long or a short labeling with14C-glucosamine, showed that benzhydrazone reduced the appearance of glycopeptides of all the size-classes, including the mannose-rich glycopeptide with an approximate molecular weight of 1500. The same per cent of label was released from both untreated and benzhydrazone-treated cells after digestion with endo-β-N-acetylglucosaminidase H, an enzyme which cleaves between the N-acetylglucosamine residues in large highmannose type oligosaccharides. This indicates that the relative amount of glycoproteins sensitive to this enzyme did not differ in the two kinds of samples. PAGE analysis confirmed that the same glycoproteins were digested in both samples. They were gA, pgC, and pgD, which therefore contain high-mannose type oligosaccharides. It is concluded that benzhydrazone hinders carbohydrate addition to herpesvirus proteins at an early step.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Abbondanza, A., Franceschi, C., Licastro, F., Serafini-Cessi, F.: Properties of a glycopeptide isolated from human Tamm-Horsfall glycoprotein. Interaction with leucoagglutinin and anti-(human Tamm-Horsfall glycoprotein) antibodies. Biochem. J.187, 525–528 (1980).

    Google Scholar 

  2. Afonso, A., Charlwood, P. A., Marshall, R. D.: Isolation and characterization of glycopeptides from digests of human Tamm-Horsfall glycoprotein. Carbohydr. Res.89, 309–320 (1981).

    Google Scholar 

  3. Arima, T., Spiro, R. G.: Studies on the carbohydrate unit of thyroglobulin. Structure of the mannose-N-acetylglucosamine unit (unit A) of the human and calf proteins. J. biol. Chem.247, 1836–1848 (1972).

    Google Scholar 

  4. Campadelli-Fiume, G., Sinibaldi-Vallebona, P., Cavrini, V., Mannini-Palenzona, A.: Selective inhibition of herpes simplex virus glycoprotein synthesis by a benz-amidinohydrazone derivative. Arch. Virol.66, 179–181 (1980).

    Google Scholar 

  5. Cavrini, V., Gatti, R., Roveri, P., Giovanninetti, G., Mannini-Palenzona, A., Baserga, M.: Antiviral compounds. XII. Synthesis andin vitro antiherpetic activity of some cyclic α- and β-diketone bis-amidinohydrazones. Eur. J. med. Chem.14, 343–346 (1979).

    Google Scholar 

  6. Cohen, G. H., Long, D., Eisenberg, R. J.: Synthesis and processing of glycoproteins gD and gC of Herpes Simplex Virus type 1. J. Virol.36, 429–439 (1980).

    Google Scholar 

  7. Eberle, R., Courtney, R. C.: gA and gB glycoproteins of herpes simplex virus type 1: two forms of a single polypeptide. J. Virol.36, 665–675 (1980).

    Google Scholar 

  8. Eisenberg, R. J., Hydrean-Stern, C., Cohen, G. H.: Structural analysis of precursor and product forms of type-common envelope glycoprotein D (CP-1 antigen) of herpes simplex virus type 1. J. Virol.31, 608–620 (1979).

    Google Scholar 

  9. Ejercito, P. L., Kieff, E. D., Roizman, B.: Characterization of herpes simplex strains differing in their effects on social behaviour of infected cells. J. gen. Virol.2, 357–364 (1968).

    Google Scholar 

  10. Fletcher, A. P., Marks, G. S., Marshall, R. D., Neuberger, A.: Carbohydrates in protein. 5. Procedures for the isolation of glycopeptides from hen's-egg albumin and their oxidation by periodate. Biochem. J.87, 265–273 (1963).

    Google Scholar 

  11. Heifetz, A., Keenan, R. W., Elbein, A. D.: Mechanism of action of tunicamycin on the UDP-GlcNac: dolichyl-phosphate GlcNAc-1-phosphate transferase. Biochemistry18, 2186–2192 (1979).

    Google Scholar 

  12. Honess, R. W., Roizman, B.: Proteins specified by herpes simplex virus. XIII. Glycosylation of viral polypeptides. J. Virol.16, 1308–1326 (1975).

    Google Scholar 

  13. Hunt, L. A., Etchison, J. R., Summers, D.: Oligosaccharide chains are trimmed during synthesis of the envelope glycoprotein of vesicular stomatitis virus. Proc. Natl. Acad. Sci. U.S.A.75, 754–758 (1978).

    Google Scholar 

  14. Ito, S., Muramatsu, T., Kobata, A.: Endo-β-N-acetylglucosaminidases acting on carbohydrate moieties of glycoproteins: purification and properties of the two enzymes with different specificities fromClostridium perfringens. Arch. Biochem. Biophys.171, 78–86 (1975).

    Google Scholar 

  15. Klenk, H. D., Rott, R.: Cotranslational and posttranslational processing of viral glycoproteins. Curr. Top. Microbiol. Immunol.90, 19–48 (1980).

    Google Scholar 

  16. Klenk, H. D., Wöllert, W., Rott, R., Scholtissek, C.: Association of influenza virus proteins with cytoplasmic fractions. Virology57, 28–41 (1974).

    Google Scholar 

  17. Lowry, O. H., Rosebrough, W. J., Farr, A. L., Randall, R. J.: Protein measurement with the folin reagent. J. biol. Chem.193, 265–275 (1951).

    Google Scholar 

  18. Nakamura, K., Compans, R. W.: Effects of glucosamine, 2-deoxyglucose, and tunicamycin on glycosylation, sulfation, and assembly of influenza viral proteins. Virology84, 303–319 (1978).

    Google Scholar 

  19. Nomoto, M., Narahashi, Y.: A proteolytic enzyme of Streptomyces griseus. I. Purification of a protease of Streptomyces griseus. J. Biochem.46, 653–667 (1959).

    Google Scholar 

  20. Norrild, B.: Immunochemistry of herpes simplex virus glycoproteins. Curr. Top. Microbiol. Immunol.90, 67–106 (1980).

    Google Scholar 

  21. Olofsson, S., Blomberg, J.: Studies on glycopeptides of herpes simplex virus infected cells. Arch. Virol.55, 293–304 (1977).

    Google Scholar 

  22. Olofsson, S., Jeansson, S., Lycke, E.: Unusual lectin-binding properties of a herpes simplex virus type 1-specific glycoprotein. J. Virol.38, 564–570 (1981).

    Google Scholar 

  23. Olofsson, S., Lycke, E.: Glucosamine metabolism of herpes simplex virus infected vcells. Inhibition of glycosylation by tunicamycin and 2-deoxy-D-glucose. Arch. Virol.65, 201–209 (1980).

    Google Scholar 

  24. Pizer, L. I., Cohen, G. H., Eisenberg, R. J.: Effect of tunicamycin on herpes simplex virus glycoproteins and infectious virus production. J. Virol.34, 142–153 (1980).

    Google Scholar 

  25. Robbins, P. W., Hubbard, S. C., Turco, J. S., Wirth, D. F.: Proposal for a common oligosaccharide intermediate in the synthesis of membrane glycoproteins. Cell12, 893–900 (1977).

    Google Scholar 

  26. Schwartz, R. T., Klenk, H. D.: Inhibition of glycosylation of influenza virus hemagglutinin. J. Virol.14, 1023–1034 (1974).

    Google Scholar 

  27. Sefton, B. M., Keegstra, K.: Glycoproteins of Sindbis virus: preliminary characterization of the oligosaccharides. J. Virol.14, 522–530 (1974).

    Google Scholar 

  28. Spear, P. G.: Membrane proteins specified by herpes simplex viruses. I. Identification of four glycoprotein precursors and their products in type 1-infected cells. J. Virol.17, 991–1008 (1976).

    Google Scholar 

  29. Staneloni, R. J., Leloir, L. F.: The biosynthetic pathway of the asparagine-linked oligosaccharides of glycoproteins. Trends biochem. Sci.4, 65–67 (1979).

    Google Scholar 

  30. Tabas, I., Schlesinger, S., Kornfeld, S.: Processing of the high mannose oligosaccharide to form complex type oligosaccharides on the newly synthesized polypeptides of the vesicular stomatitis virus G protein and the IgG heavy chain. J. biol. Chem.253, 716–722 (1978).

    Google Scholar 

  31. Takatsuki, A., Tamura, G.: Effect of tunicamycin on the synthesis of macromolecules in cultures of chick embryo fibroblasts infected with Newcastle disease virus. J. Antib. (Tokyo)24, 785–794 (1971).

    Google Scholar 

  32. Tarentino, A. L., Maley, F.: Purification and properties of an endo-β-N-acetylglucosaminidase fromStreptomyces griseus. J. biol. chem.249, 811–816 (1974).

    Google Scholar 

  33. Tkacz, J. S., Lampen, J. O.: Tunicamycin inhibition of polyisoprenil N-acetylglucosaminyl pyrophosphate formation in calf liver microsomes. Biochem. biophys. Res. Commun.65, 248–257 (1975).

    Google Scholar 

  34. Warren, L.: The thiobarbituric acid assay of sialic acids. J. biol. Chem.234, 1971–1975 (1959).

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Istituto di Patologia Generale, Università di Bologna, Bologna, Italy

    F. Serafini-Cessi

  2. Istituto di Microbiologia e Virologia, Università di Bologna, Bologna, Italy

    G. Campadelli-Fiume

Authors
  1. F. Serafini-Cessi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. G. Campadelli-Fiume
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

With 5 Figures

Rights and permissions

Reprints and permissions

About this article

Cite this article

Serafini-Cessi, F., Campadelli-Fiume, G. Studies on benzhydrazone, a specific inhibitor of herpesvirus glycoprotein synthesis. Size distribution of glycopeptides and endo-β-N-acetylglucosaminidase-H treatment. Archives of Virology 70, 331–343 (1981). https://doi.org/10.1007/BF01320248

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01320248

Keywords

Search

Navigation