Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1984 Sep;51(3):838–844. doi: 10.1128/jvi.51.3.838-844.1984

Processing of N-linked oligosaccharides from precursor- to mature-form herpes simplex virus type 1 glycoprotein gC.

F Serafini-Cessi, F Dall'Olio, L Pereira, G Campadelli-Fiume
PMCID: PMC255852  PMID: 6088806

Abstract

Immature and mature forms of glycoprotein gC were purified by immunoadsorbent from herpes simplex virus type 1-infected BHK cells labeled with [3H]mannose for a 20-min pulse or for 11 h followed by a 3-h chase. The nature of N-asparagine-linked oligosaccharides carried by the immature form, pgC (molecular weight = 92,000), and the mature gC (molecular weight = 120,000) has been investigated. All pronase-digested glycopeptides of pgC were susceptible to endo-beta-N-acetylglucosaminidase H treatment; thus they have a high-mannose structure. Using thin-layer chromatography to separate endo-beta-N-acetylglucosaminidase H-cleaved oligosaccharides, polymannosyl chains of different sizes, ranging from Man9GlcNAc to Man5GlcNAc, were separated. The major components were Man8GlcNAc and Man7GlcNAc, suggesting that pgC labeled in a 20-min pulse represents the form of glycoprotein already routed to the Golgi apparatus. Analysis of glycopeptides of mature gC showed that the majority (95%) of N-linked glycans were converted to complex-type glycans. Ion-exchange chromatography and affinity chromatography on concanavalin A-Sepharose and leucoagglutinin-agarose revealed that diantennary and triantennary glycans predominated, whereas tetrantennary chains were not present. Parts of the di- and triantennary chains were not fully sialylated. The high heterogeneity of complex-type chains found in mature gC may be related to the high number of N-glycosylation sites of the glycoprotein as predicted by DNA sequencing studies (Frink et al., J. Virol. 45:634-647, 1983).

Full text

PDF
838

Images in this article

839Image
on p.839
841Image
on p.841

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  2. Burke D., Keegstra K. Carbohydrate structure of Sindbis virus glycoprotein E2 from virus grown in hamster and chicken cells. J Virol. 1979 Feb;29(2):546–554. doi: 10.1128/jvi.29.2.546-554.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Campadelli-Fiume G., Poletti L., Dall'Olio F., Serafini-Cessi F. Infectivity and glycoprotein processing of herpes simplex virus type 1 grown in a ricin-resistant cell line deficient in N-acetylglucosaminyl transferase I. J Virol. 1982 Sep;43(3):1061–1071. doi: 10.1128/jvi.43.3.1061-1071.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cohen G. H., Long D., Eisenberg R. J. Synthesis and processing of glycoproteins gD and gC of herpes simplex virus type 1. J Virol. 1980 Nov;36(2):429–439. doi: 10.1128/jvi.36.2.429-439.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cummings R. D., Kornfeld S. Characterization of the structural determinants required for the high affinity interaction of asparagine-linked oligosaccharides with immobilized Phaseolus vulgaris leukoagglutinating and erythroagglutinating lectins. J Biol Chem. 1982 Oct 10;257(19):11230–11234. [PubMed] [Google Scholar]
  6. Eberle R., Courtney R. J. gA and gB glycoproteins of herpes simplex virus type 1: two forms of a single polypeptide. J Virol. 1980 Dec;36(3):665–675. doi: 10.1128/jvi.36.3.665-675.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Eisenberg R. J., Ponce de Leon M., Pereira L., Long D., Cohen G. H. Purification of glycoprotein gD of herpes simplex virus types 1 and 2 by use of monoclonal antibody. J Virol. 1982 Mar;41(3):1099–1104. doi: 10.1128/jvi.41.3.1099-1104.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Frink R. J., Eisenberg R., Cohen G., Wagner E. K. Detailed analysis of the portion of the herpes simplex virus type 1 genome encoding glycoprotein C. J Virol. 1983 Feb;45(2):634–647. doi: 10.1128/jvi.45.2.634-647.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Godelaine D., Spiro M. J., Spiro R. G. Processing of the carbohydrate units of thyroglobulin. J Biol Chem. 1981 Oct 10;256(19):10161–10168. [PubMed] [Google Scholar]
  10. Hammarström S., Hammarström M. L., Sundblad G., Arnarp J., Lönngren J. Mitogenic leukoagglutinin from Phaseolus vulgaris binds to a pentasaccharide unit in N-acetyllactosamine-type glycoprotein glycans. Proc Natl Acad Sci U S A. 1982 Mar;79(5):1611–1615. doi: 10.1073/pnas.79.5.1611. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Heine J. W., Spear P. G., Roizman B. Proteins specified by herpes simplex virus. VI. Viral proteins in the plasma membrane. J Virol. 1972 Mar;9(3):431–439. doi: 10.1128/jvi.9.3.431-439.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Honess R. W., Roizman B. Proteins specified by herpes simplex virus. XIII. Glycosylation of viral polypeptides. J Virol. 1975 Nov;16(5):1308–1326. doi: 10.1128/jvi.16.5.1308-1326.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hsieh P., Rosner M. R., Robbins P. W. Selective cleavage by endo-beta-N-acetylglucosaminidase H at individual glycosylation sites of Sindbis virion envelope glycoproteins. J Biol Chem. 1983 Feb 25;258(4):2555–2561. [PubMed] [Google Scholar]
  14. Hubbard S. C., Ivatt R. J. Synthesis and processing of asparagine-linked oligosaccharides. Annu Rev Biochem. 1981;50:555–583. doi: 10.1146/annurev.bi.50.070181.003011. [DOI] [PubMed] [Google Scholar]
  15. Johnson D. C., Spear P. G. Monensin inhibits the processing of herpes simplex virus glycoproteins, their transport to the cell surface, and the egress of virions from infected cells. J Virol. 1982 Sep;43(3):1102–1112. doi: 10.1128/jvi.43.3.1102-1112.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Johnson D. C., Spear P. G. O-linked oligosaccharides are acquired by herpes simplex virus glycoproteins in the Golgi apparatus. Cell. 1983 Mar;32(3):987–997. doi: 10.1016/0092-8674(83)90083-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Klenk H. D., Keil W., Niemann H., Geyer R., Schwarz R. T. The characterization of influenza A viruses by carbohydrate analysis. Curr Top Microbiol Immunol. 1983;104:247–257. doi: 10.1007/978-3-642-68949-9_15. [DOI] [PubMed] [Google Scholar]
  18. Kobata A. Use of endo- and exoglycosidases for structural studies of glycoconjugates. Anal Biochem. 1979 Nov 15;100(1):1–14. doi: 10.1016/0003-2697(79)90102-7. [DOI] [PubMed] [Google Scholar]
  19. Krusius T., Finne J., Rauvala H. The structural basis of the different affinities of two types of acidic N-glycosidic glycopeptides for concanavalin A--sepharose. FEBS Lett. 1976 Nov 15;72(1):117–120. doi: 10.1016/0014-5793(76)80911-8. [DOI] [PubMed] [Google Scholar]
  20. Kumarasamy R., Blough H. A. Characterization of oligosaccharides of highly purified glycoprotein gC of herpes simplex virus type 1 (HSV-1). Biochem Biophys Res Commun. 1982 Dec 31;109(4):1108–1115. doi: 10.1016/0006-291x(82)91891-5. [DOI] [PubMed] [Google Scholar]
  21. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  22. Marsden H. S., Buckmaster A., Palfreyman J. W., Hope R. G., Minson A. C. Characterization of the 92,000-dalton glycoprotein induced by herpes simplex virus type 2. J Virol. 1984 May;50(2):547–554. doi: 10.1128/jvi.50.2.547-554.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Olofsson S., Blomberg J., Lycke E. O-glycosidic carbohydrate-peptide linkages of Herpes simplex virus glycoproteins. Arch Virol. 1981;70(4):321–329. doi: 10.1007/BF01320247. [DOI] [PubMed] [Google Scholar]
  24. Olofsson S., Norrild B., Andersen A. B., Pereira L., Jeansson S., Lycke E. Populations of herpes simplex virus glycoprotein gC with and without affinity for the N-acetyl-galactosamine specific lectin of Helix pomatia. Arch Virol. 1983;76(1):25–38. doi: 10.1007/BF01315701. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Palfreyman J. W., Haarr L., Cross A., Hope R. G., Marsden H. S. Processing of herpes simplex virus type 1 glycoproteins: two-dimensional gel analysis using monoclonal antibodies. J Gen Virol. 1983 Apr;64(Pt 4):873–886. doi: 10.1099/0022-1317-64-4-873. [DOI] [PubMed] [Google Scholar]
  26. Peake M. L., Nystrom P., Pizer L. I. Herpesvirus glycoprotein synthesis and insertion into plasma membranes. J Virol. 1982 May;42(2):678–690. doi: 10.1128/jvi.42.2.678-690.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Pereira L., Klassen T., Baringer J. R. Type-common and type-specific monoclonal antibody to herpes simplex virus type 1. Infect Immun. 1980 Aug;29(2):724–732. doi: 10.1128/iai.29.2.724-732.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Pizer L. I., Cohen G. H., Eisenberg R. J. Effect of tunicamycin on herpes simplex virus glycoproteins and infectious virus production. J Virol. 1980 Apr;34(1):142–153. doi: 10.1128/jvi.34.1.142-153.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Rush J. S., Turco S. J., Laine R. A. Erythroglycan biosynthesis in K-562 cells. Inhibition of synthesis by tunicamycin and lack of attachment to the G-protein of vesicular-stomatitis virus. Biochem J. 1981 Jan 1;193(1):361–365. doi: 10.1042/bj1930361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Serafini-Cessi F., Campadelli-Fiume G. Studies on benzhydrazone, a specific inhibitor of herpesvirus glycoprotein synthesis. Size distribution of glycopeptides and endo-beta-N-acetylglucosaminidase-H treatment. Arch Virol. 1981;70(4):331–343. doi: 10.1007/BF01320248. [DOI] [PubMed] [Google Scholar]
  31. Serafini-Cessi F., Dall'Olio F. Guinea-pig kidney beta-N-acetylgalactosaminyltransferase towards Tamm-Horsfall glycoprotein. Requirement of sialic acid in the acceptor for transferase activity. Biochem J. 1983 Dec 1;215(3):483–489. doi: 10.1042/bj2150483. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Serafini-Cessi F., Dall'Olio F., Scannavini M., Campadelli-Fiume G. Processing of herpes simplex virus-1 glycans in cells defective in glycosyl transferases of the Golgi system: relationship to cell fusion and virion egress. Virology. 1983 Nov;131(1):59–70. doi: 10.1016/0042-6822(83)90533-0. [DOI] [PubMed] [Google Scholar]
  33. Serafini-Cessi F., Franceschi C., Sperti S. Specific interaction of human Tamm-Horsfall gylcoprotein with leucoagglutinin, a lectin from Phaseolus vulgaris (red kidney bean). Biochem J. 1979 Nov 1;183(2):381–388. doi: 10.1042/bj1830381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. 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. 1976 Mar;17(3):991–1008. doi: 10.1128/jvi.17.3.991-1008.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Spear P. G., Roizman B. Proteins specified by herpes simplex virus. V. Purification and structural proteins of the herpesvirion. J Virol. 1972 Jan;9(1):143–159. doi: 10.1128/jvi.9.1.143-159.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Spiro R. G., Spiro M. J. Studies on the synthesis and processing of the asparagine-linked carbohydrate units of glycoproteins. Philos Trans R Soc Lond B Biol Sci. 1982 Dec 24;300(1099):117–127. doi: 10.1098/rstb.1982.0160. [DOI] [PubMed] [Google Scholar]
  37. Tabas I., Schlesinger S., Kornfeld S. Processing of high mannose oligosaccharides 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. 1978 Feb 10;253(3):716–722. [PubMed] [Google Scholar]
  38. Tarentino A. L., Maley F. Purification and properties of an endo-beta-N-acetylglucosaminidase from Streptomyces griseus. J Biol Chem. 1974 Feb 10;249(3):811–817. [PubMed] [Google Scholar]
  39. Wenske E. A., Bratton M. W., Courtney R. J. Endo-beta-N-acetylglucosaminidase H sensitivity of precursors to herpes simplex virus type 1 glycoproteins gB and gC. J Virol. 1982 Oct;44(1):241–248. doi: 10.1128/jvi.44.1.241-248.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES