Skip to main content
SpringerLink
Log in

Intracellular processing and antigenic maturation of measles virus hemagglutinin protein

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

Summary

The intracellular processing and antigenic maturation of the measles virus (MV) hemagglutinin (H) protein in virus infected cells were probed with murine monoclonal antibodies (Mabs) that reacted with continuous and discontinuous epitopes. The antibodies distinguished between the immature, cotranslational monomeric form of the protein and the mature, dimeric hemagglutinin structure. This was evidenced by testing of immunoreactivity of the Mabs with synthetic peptides, by in vitro synthesized H protein analysis, and by pulse-chase analysis of gel separated monomeric and dimeric forms of the H protein. Time kinetics analysis showed that the protein was synthesized as monomers and most of them were converted into dimers witht 1/2 about 30 min. The H protein remained endoglycosidase H (Endo H) sensitive up to 30 min and started to acquire partial resistance to Endo H between 30 and 60 min (t 1/2 about 60 min) after synthesis. Oligomerization of the H protein was unaffected in virus infected cells treated with a compound (carbonylcyanide m-chlorophenylhydrazone, CCCP) that blocks transport from the endoplasmic reticulum (ER) to the Golgi complex. These results suggest that the H protein dimerization takes place in the ER before its transport to the medial Golgi complex. The Mabs specific for discontinuous epitopes reacted with the H protein in cells treated with CCCP. Thus conformational antigenic epitope formation appears to take place in the ER.

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. Åkerlind-Stopner B, Utter G, Mufson MA, Örell C, Lerner RA, Norrby E (1990) A subgroup-specific antigenic site in the G protein of respiratory syncytial virus forms a disulfide-bonded loop. J Virol 64: 5143–5148

    Google Scholar 

  2. Alkhatib G, Briedis DJ (1986) The predicted primary structure of the measles virus hemagglutinin. Virology 150: 479–490

    Google Scholar 

  3. Boulay F, Doms RW, Webster RG, Helenius A (1988) Posttranslational oligomerization and cooperative acid activation of mixed influenza hemagglutinin trimers. J Cell Biol 106: 629–639

    Google Scholar 

  4. Carter MJ, Willcocks MM, Löffer S, ter Meulen V (1982) Relationship between monoclonal antibody biding sites on the measles virus haemagglutinin. J Gen Virol 63: 113–120

    Google Scholar 

  5. Copeland CS, Doms RW, Bolzau EM, Webster RG, Helenius A (1986) Assembly of influenza hemagglutinin trimers and its role in intracellular transport. J Cell Biol 103: 1179–1191

    Google Scholar 

  6. Copeland CS, Zimmer KP, Wagner KR, Healey GA, Mellman I, Helenius A (1988) Folding, trimerization, and transport are sequential events in the biogenesis of influenza virus hemagglutinin. Cell 53: 197–209

    Google Scholar 

  7. Doms RW, Keller DS, Helenius A, Balch WE (1987) Role of triphosphate in regulating the assembly and transport of vesicular stomatitits virus G protein trimers. J Cell Biol 105: 1957–1969

    Google Scholar 

  8. Doms RW, Ruusala A, Machamer CM, Helenius A, Rose JK (1988) Differential effects of mutations in three domains on folding, quaternary structure, and intracellular transport of vesicular stomatitis virus G protein. J Cell Biol 107: 89–99

    Google Scholar 

  9. Doms RW, Lamb RA, Rose JK, Helenius A (1993) Minireview: folding and assembly of viral membrane proteins. Virology 193: 545–562

    Google Scholar 

  10. Eschle D (1988) Diploma thesis. University of Zurich, Zurich, Switzerland

    Google Scholar 

  11. Freeman RB (1984) Native disulphide bond formation in protein biosynthesis: evidence for the role of protein disulphide isomerase. Trends Biochem Sci 9: 438–441

    Google Scholar 

  12. Fries E, Rothman JE (1980) Transport of vesicular stomatitis virus glycoprotein in a cell-free extract. Proc Natl Acad Sci USA 77: 3870–3874

    Google Scholar 

  13. Gething MJ, Mclammon K, Sambrook J (1986) Expression of wild type and mutant forms of influenza hemagglutinin: the role of folding in intracellular transport. Cell 46: 939–950

    Google Scholar 

  14. Gombart AF, Hirano A, Wong TC (1993) Conformational maturation of measles virus nucleocapsid protein. J Virol 67: 4133–4141

    Google Scholar 

  15. Houghten RA (1985) General method for the rapid solid phase synthesis of large numbers of peptides: specificity of antigenantibody interaction at the level of individual amino acids. Proc Natl Acad Sci USA 82: 5131–5135

    Google Scholar 

  16. Houghten RA, Bray MK, DeGraw ST, Kirby CJ (1986) Simplified procedure for carrying out simultaneous multiple hydrogen fluoride cleavage of protected peptide resins. Int J Peptide Protein Res 27: 675–680

    Google Scholar 

  17. Hu A, Sheshberadaran H, Norrby E, Kövamees J (1993) Molecular characterization of epitopes on the measles virus hemagglutinin protein. Virology 192: 351–354

    Google Scholar 

  18. Hu A, Cattaneo R, Schwartz S, Norrby E (1994) Role of N-linked oligosaccharide chains in the processing and antigenicity of the measles virus hemagglutinin protein. J Gen Virol (in press)

  19. Kornfeld R, Kornfeld S (1985) Assembly of asparagine-linked oligosaccharides. Annu Rev Biochem 54: 631–664

    Google Scholar 

  20. Kreis TE, Lodish HF (1986) Oligomerization is essential for transport of vesicular stomatitis viral glycoprotein to the cell surface. Cell 46: 929–937

    Google Scholar 

  21. Mäkelä MJ, Lund GA, Salmi AA (1989a) Antigenicity of the measles virus haemagglutinin studied by using synthetic peptides. J Gen Virol 70: 603–614

    Google Scholar 

  22. Mäkelä MJ, Salmi AA, Norrby E, Wild TF (1989b) Monoclonal antibodies against measles virus haemagglutinin react with synthetic peptides. Scand J Immunol 30: 225–231

    Google Scholar 

  23. Mottet G, Portner A, Roux L (1986) Drastic immunoreactivity changes between the immature and mature forms of the Sendai virus HN and F0 glycoproteins. J Virol 59: 132–141

    Google Scholar 

  24. Ng DT, Randall RE, Lamb RA (1989) Intracellular maturation and transport of the SV5 hemagglutinin-neuraminidase: specific and transient association with GRP-Bip in the endoplasmic reticulum and extensive internalization from cell surface. J Cell Biol 109: 3273–3289

    Google Scholar 

  25. Norrby E, Hammarskjöld B (1972) Structural components of measles virus. Microbios 5: 17–29

    Google Scholar 

  26. Norrby E, Chen SN, Togashi T, Sheshberadaran H, Johnson KP (1982) Five measles virus antigens demonstrated by use of mouse hybridoma antibodies in productively infected tissue cells. Arch Virol 71: 1–11

    Google Scholar 

  27. Norrby E, Oxman M (1990) Measles virus. In: Fields BN, Knipe DM (eds) Virology, 2nd edn. Raven Press, New York, pp 1013–1044

    Google Scholar 

  28. Pedersen IR, Bog-Hansen TC, Dalsgaord K, Heegaord PMH (1992) Iscom immunization with synthetic peptides representing measles virus hemagglutinin. Virus Res 24: 145–159

    Google Scholar 

  29. Peters J Jr, Davison LK (1982) The biosynthesis of rate serum albumin: in vivo studies on the formation of disulfide bonds. J Biol Chem 257: 8847–8853

    Google Scholar 

  30. Rota JS, Hummel KB, Rota PA, Bellini WJ (1992) Genetic variation of the glycoprotein genes of current wild-type measles isolates. Virology 188: 135–142

    Google Scholar 

  31. Sheshberadaran H, Chen SN, Norrby E (1983) Monoclonal antibodies against five structural components of measles virus. I. Characterization of antigenic determinants on nine strains of measles virus. Virology 128: 341–353

    Google Scholar 

  32. Sheshberadaran H, Norrby E (1986) Characterization of epitopes on the measles virus hemagglutinin protein. Virology 152: 58–65

    Google Scholar 

  33. Sheshberadaran H, Payne GL (1988) Protein antigen-monoclonal antibody contact sites investigated by limited proteolysis of monoclonal antibody-bound antigen: protein “footprinting”. Proc Natl Acad Sci USA 85: 1–5

    Google Scholar 

  34. Tartakoff AM (1986) Temperature and energy dependence of secretory protein transport in the exocrine pancreas. EMBO J 5: 1477–1482

    Google Scholar 

  35. Vidal S, Mottet G, Kolakofsky D, Roux L (1989) Addition of high-mannose sugars must precede disulfide bond formation for proper folding of Sendai virus glycoproteins. J Virol 63: 892–900

    Google Scholar 

  36. Waxham MN, Merz DV, Wolinsky JS (1986) Intracellular maturation of mumps virus hemagglutinin-neuraminidase glycoprotein: conformational changes detected with monoclonal antibodies. J Virol 59: 392–400

    Google Scholar 

  37. Yamada A, Takeuchi K, Hishiyama M (1988) Intracellular processing of mumps virus glycoproteins. Virology 165: 268–273

    Google Scholar 

  38. Yewdell JW, Yellen A, Bachi T (1988) Monoclonal antibodies localize events in the folding, assembly, and intracellular transport of the influenza virus hemagglutinin glycoprotein. Cell 52: 843–852

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Virology, School of Medicine, Karolinska Institute, Stockholm, Sweden

    A. Hu, J. Kövamees & E. Norrby

Authors
  1. A. Hu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J. Kövamees
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. E. Norrby
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hu, A., Kövamees, J. & Norrby, E. Intracellular processing and antigenic maturation of measles virus hemagglutinin protein. Archives of Virology 136, 239–253 (1994). https://doi.org/10.1007/BF01321055

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01321055

Keywords

Search

Navigation