Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1979 Nov;76(11):5621–5625. doi: 10.1073/pnas.76.11.5621

Glycoproteins of Sendai virus are transmembrane proteins.

D S Lyles
PMCID: PMC411701  PMID: 230485

Abstract

Radiolabeled Sendai viral envelope proteins were incorporated into human erythrocyte membranes by the process of fusion of the viral envelope with the erythrocyte membrane. Inside-out (IO) vesicles were prepared from the erythrocyte membranes containing viral proteins, and the presence of viral proteins was assessed by electrophoresis in sodium dodecyl sulfate/polyacrylamide gels. Proteolysis with trypsin, chymotrypsin, or Pronase, which digests only the external surface of the IO vesicles (the cytoplasmic surface of the erythrocyte membrane) revealed that the viral nucleocapsid and the nonglycosylated inner-envelope (M) proteins were present on the external surface. In addition, small segments of the viral envelope glycoproteins (HN and F1) were removed by these proteases, while the major portions of the glycoproteins were protected from digestion, indicating that in the erythrocyte membrane they are transmembrane proteins. Results of experiments carried out on right side-out membranes, unsealed membranes, and membranes containing attached virus that was unable to fuse indicated that the results obtained with IO membranes could not be accounted for by contamination with these membrane species. The identify of the proteolysis products was confirmed by peptide mapping. The selective exposure of the cytoplasmic surface, and hence the internal components of the virus, in IO vesicles makes this membrane system an attractive model for studying the interactions involved in virus maturation at host cell membranes.

Full text

PDF
5621

Images in this article

5622Image
on p.5622
5622Image
on p.5622
5623Image
on p.5623
5623Image
on p.5623
5624Image
on p.5624

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. Bächi T., Aguet M., Howe C. Fusion of erythrocytes by Sendai virus studied by immuno-freeze-etching. J Virol. 1973 Jun;11(6):1004–1012. doi: 10.1128/jvi.11.6.1004-1012.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chen C., Compans R. W., Choppin P. W. Parainfluenza virus surface projections: glycoproteins with haemagglutinin and neuraminidase activities. J Gen Virol. 1971 Apr;11(1):53–58. doi: 10.1099/0022-1317-11-1-53. [DOI] [PubMed] [Google Scholar]
  4. Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed] [Google Scholar]
  5. Dales S. Early events in cell-animal virus interactions. Bacteriol Rev. 1973 Jun;37(2):103–135. doi: 10.1128/br.37.2.103-135.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Homma M., Ouchi M. Trypsin action on the growth of Sendai virus in tissue culture cells. 3. Structural difference of Sendai viruses grown in eggs and tissue culture cells. J Virol. 1973 Dec;12(6):1457–1465. doi: 10.1128/jvi.12.6.1457-1465.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Howe C., Morgan C. Interactions between Sendai virus and human erythrocytes. J Virol. 1969 Jan;3(1):70–81. doi: 10.1128/jvi.3.1.70-81.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Katz F. N., Rothman J. E., Lingappa V. R., Blobel G., Lodish H. F. Membrane assembly in vitro: synthesis, glycosylation, and asymmetric insertion of a transmembrane protein. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3278–3282. doi: 10.1073/pnas.74.8.3278. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  11. Lamb R. A., Choppin P. W. Determination by peptide mapping of the unique polypeptides in Sendai virions and infected cells. Virology. 1978 Feb;84(2):469–478. doi: 10.1016/0042-6822(78)90263-5. [DOI] [PubMed] [Google Scholar]
  12. Lamb R. A., Choppin P. W. The synthesis of Sendai virus polypeptides in infected cells. III. Phosphorylation of polypeptides. Virology. 1977 Sep;81(2):382–397. doi: 10.1016/0042-6822(77)90154-4. [DOI] [PubMed] [Google Scholar]
  13. Lamb R. A., Mahy B. W., Choppin P. W. The synthesis of sendai virus polypeptides in infected cells. Virology. 1976 Jan;69(1):116–131. doi: 10.1016/0042-6822(76)90199-9. [DOI] [PubMed] [Google Scholar]
  14. Mountcastle W. E., Compans R. W., Caliguiri L. A., Choppin P. W. Nucleocapsid protein subunits of simian virus 5, Newcastle disease virus, and Sendai virus. J Virol. 1970 Nov;6(5):677–684. doi: 10.1128/jvi.6.5.677-684.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Scheid A., Caliguiri L. A., Compans R. W., Choppin P. W. Isolation of paramyxovirus glycoproteins. Association of both hemagglutinating and neuraminidase activities with the larger SV5 glycoprotein. Virology. 1972 Dec;50(3):640–652. doi: 10.1016/0042-6822(72)90418-7. [DOI] [PubMed] [Google Scholar]
  16. Scheid A., Choppin P. W. Identification of biological activities of paramyxovirus glycoproteins. Activation of cell fusion, hemolysis, and infectivity of proteolytic cleavage of an inactive precursor protein of Sendai virus. Virology. 1974 Feb;57(2):475–490. doi: 10.1016/0042-6822(74)90187-1. [DOI] [PubMed] [Google Scholar]
  17. Scheid A., Choppin P. W. Isolation and purification of the envelope proteins of Newcastle disease virus. J Virol. 1973 Feb;11(2):263–271. doi: 10.1128/jvi.11.2.263-271.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Scheid A., Choppin P. W. Protease activation mutants of sendai virus. Activation of biological properties by specific proteases. Virology. 1976 Jan;69(1):265–277. doi: 10.1016/0042-6822(76)90213-0. [DOI] [PubMed] [Google Scholar]
  19. Scheid A., Choppin P. W. Two disulfide-linked polypeptide chains constitute the active F protein of paramyxoviruses. Virology. 1977 Jul 1;80(1):54–66. doi: 10.1016/0042-6822(77)90380-4. [DOI] [PubMed] [Google Scholar]
  20. Shimizu K., Isida N. The smallest protein of Sendi virus: its candidate function of binding nucleocaspsid to envelope. Virology. 1975 Oct;67(2):427–437. [PubMed] [Google Scholar]
  21. Steck T. L. The organization of proteins in the human red blood cell membrane. A review. J Cell Biol. 1974 Jul;62(1):1–19. doi: 10.1083/jcb.62.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Tozawa H., Watanabe M., Ishida N. Structural components of Sendai virus. Serological and physicochemical characterization of hemagglutinin subunit associated with neuraminidase activity. Virology. 1973 Sep;55(1):242–253. doi: 10.1016/s0042-6822(73)81027-x. [DOI] [PubMed] [Google Scholar]
  23. Wirth D. F., Katz F., Small B., Lodish H. F. How a single Sindbis virus mRNA directs the synthesis of one soluble protein and two integral membrane glycoproteins. Cell. 1977 Feb;10(2):253–263. doi: 10.1016/0092-8674(77)90219-7. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES