1887

Abstract

The envelope glycoprotein, gpl60, of human (HIV) and simian (SIV) immunodeficiency viruses mediates virus- host cell binding followed by fusion of the viral and plasma membranes. The envelope proteins are known to exist as non-covalently associated oligomers on the virus surface. The production of permanent mammalian cell lines that constitutively secrete relatively high levels of soluble forms of SIV gpl60 is described and we show that these proteins are secreted predominantly as tetramers with lower levels of dimer forms. Oligomeric forms were purified to greater than 90 % purity using a simple gel filtration method. The purified proteins bind CD4 suggesting that they remain in their native conformation. The purified oligomeric proteins provide the basis for more relevant structural, functional and immunological studies than recombinant gp120 as they more closely resemble the envelope protein oligomer.

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1994-01-01
2024-04-19
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References

  1. Banda N. K., Bernier J., Kurahara D. K., Kurrle R., Haigwood N., Sekaly R.-P., Finkel T. H. 1992; Crosslinking CD4 by human immunodeficiency virus gp120 primes T cells for activation-induced apoptosis. Journal of Experimental Medicine 176:1099–1106
    [Google Scholar]
  2. Berman P. W., Riddle L., Nakamura G., Haffar O. K., Nunes W. M., Skehel P., Bryn R., Groopman J., Metthews T., Gregory T. 1989; Expression and immunogenicity of the extracellular domain of the human immunodeficiency virus type 1 envelope glycoprotein, gp160. Journal of Virology 63:3489–3498
    [Google Scholar]
  3. Chakrabarti S., Mizukami T., Franchini G., Moss B. 1990; Synthesis, oligomerization, and biological activity of the human immunodeficiency virus type 2 envelope glycoprotein expressed by recombinant vaccinia virus. Virology 178:134–142
    [Google Scholar]
  4. Chapman B. S., Thayer R. M., Vincent K. A., Haigwood N. I. 1991; Effect of intron A from human cytomegalovirus (Towne) immediate early gene on heterologous expression in mammalian cells. Nucleic Acids Research 19:3979–3986
    [Google Scholar]
  5. Cockett M. I., Bebbington C. R., Yarranton G. 1991; The use of engineered E1A genes to transactivate the hCMV-MIE promoter in permanent CHO ceil lines. Nucleic Acids Research 19:319–325
    [Google Scholar]
  6. Doms R. W., Earl P. L., Chakrabarti S., Moss B. 1990; Human immunodeficiency virus types 1 and 2 and simian immunodeficiency virus env proteins possess a functionally conserved assembly domain. Journal of Virology 64:3537–3540
    [Google Scholar]
  7. Doms R. W., Earl P. L., Moss B. 1991; The assembly of the HIV-1 env glycoprotein into dimers and tetramers. In Mechanisms and Specificity of HIV Entry into Host Cells pp. 203–221 Düzgüneş N. Edited by New York: Plenum Press;
    [Google Scholar]
  8. Dunbar B. S., Schwoebel E. D. 1990; Preparation of polyclonal antibodies. In Guide to Protein Purification pp. 663–670 Deutscher M. P. Edited by New York: Academic Press;
    [Google Scholar]
  9. Earl P. L., Doms R. W., Moss B. 1990; Oligomeric structure of the human immunodeficiency virus type 1 envelope glycoprotein. Proceedings of the National Academy of Sciences, U.S.A 87:648–652
    [Google Scholar]
  10. Earl P. L., Doms R. W., Moss B. 1991; Folding, interaction with GRP78-BiP, assembly, and transport of the human immunodeficiency virus type 1 envelope protein. Journal of Virology 65:2047–2055
    [Google Scholar]
  11. Earl P. L., Doms R. W., Moss B. 1992; Multimeric CD4 binding exhibited by human and simian immunodeficiency virus envelope protein dimers. Journal of Virology 66:5610–5614
    [Google Scholar]
  12. Harris T. J. R., Patel T., Marston F. A. O., Little S., Emtage J. S. 1986; Cloning of cDNA coding for human tissue-type plasminogen activator and its expression in Escherichia coli. Molecular Biology and Medicine 3:279–292
    [Google Scholar]
  13. Kent K. A., Gritz L., Stalland G., Cranage M., Collignon C., Thiriart C., Corcoran T., Silvera P., Stott E. J. 1991; Production of monoclonal antibodies to simian immunodeficiency virus envelope glycoproteins. AIDS 5:829–836
    [Google Scholar]
  14. Kieny M. P., Lathe R., Riviere Y., Dott K., Schmitt D., Girard M., Montagnier L., Lecocq J.-P. 1988; Improved antigenicity of the HIV env protein by cleavage site removal. Protein Engineering 2:219–225
    [Google Scholar]
  15. Kingston R. E., Kaufman R. J., Bebbington C. R., Rolfe M. R. 1992; Amplification using CHO cell expression vectors. In Short Protocols in Molecular Biology, 2nd edn. pp. 55–62 Ausubel F. M., Brent R., Kingston R. E., Moore D. D., Seidman J. D., Smith J. A., Struhl K. Edited by New York: John Wiley;
    [Google Scholar]
  16. Kowalski M., Potz J., Basiripour L., Dorfman T., Goh W. C., Terwilliger E., Dayton A., Rosen C., Haseltine W., Sodroski J. 1987; Functional regions of the envelope glycoprotein of human immunodeficiency virus type 1. Science 237:1351–1355
    [Google Scholar]
  17. Mcclure M. O., Marsh M., Weiss R. A. 1988; Human immunodeficiency virus infection of CD4-bearing cells occurs by a pH-independent mechanism. EMBO Journal 7:513–518
    [Google Scholar]
  18. Mccune J. M., Rabin L. B., Feinburg M. B., Lieberman M., Koersk J. C., Reyes G. R., Weissman I. L. 1988; Endo-proteolytic cleavage of gp160 is required for the activation of human immunodeficiency virus. Cell 53:55–67
    [Google Scholar]
  19. Mcdougal J. S., Nicholson J. K. A., Cross G. D., Cort S. P., Kennedy M. S., Mawle A. C. 1986; Binding of the human retrovirus HTLV-III/LAV/ARV/HIV to the CD4 (T4) molecule: conformation dependence, epitope mapping, antibody inhibition and potential for idiotypic mimicry. Journal of Immunology 137:2937–2944
    [Google Scholar]
  20. Moore J. P. 1990; Simple methods for monitoring HIV-1 and HIV-2 gp120 binding to soluble CD4 by enzyme-linked immunosorbent assay: HIV-2 has a 25-fold lower affinity than HIV-1 for soluble CD4. AIDS 4:297–305
    [Google Scholar]
  21. Pinter A., Honnen W. J., Tilley S. A., Bona C., Zaghouani H., Gorny M. K., Zolla-Pazner S. 1989; Oligomeric structure of gp41, the transmembrane protein of human immunodeficiency virus type 1. Journal of Virology 63:2674–2679
    [Google Scholar]
  22. Plannelles V., Haigwood N. L., Marthas M. L., Mann K. A., Scandella C., Lidster W. D., Shuster J. R., Van Kuyk R., Marx P. A., Gardner M. B., Luciw P. A. 1991; Functional and immunological characterization of SIV envelope glycoprotein produced in genetically engineered mammalian cells. AIDS Research and Human Retroviruses 7:889–898
    [Google Scholar]
  23. Putney S., Matthews T. J., Robey W., Lynn D., Robert-Guroff M., Mueller W., Langlois A., Ghrayeb J., Petteway S., Weinhold K., Fischinger P., Wong-Staal F., Gallo R., Bolognbsi D. 1986; HTLV III/LAV neutralising antibodies to an E. coli-produced fragment of the virus envelope. Science 234:1392–1395
    [Google Scholar]
  24. Rey M.-A., Laurent A. G., Mcclure J., Krust B., Montagnier L., Hovanessian A. G. 1990; Transmembrane envelope glycoprotein of human immunodeficiency virus type II and simian immunodeficiency virus SIV-mac exist as homodimers. Journal of Virology 64:922–926
    [Google Scholar]
  25. Rud E. W., Yon J. R., Larder B. A., Clarke B. E., Cook N., Cranage M. P. 1992; Infectious molecular clones of SIVmac32H; Nef deletion controls ability to reisolate virus from rhesus macaques. In Vaccines 92: Modem Approaches to New Vaccines Including Prevention of AIDS pp. 229–235 Brown F., Chanock R. M., Ginsberg H. S., Lerner R. A. Edited by New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  26. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: A Laboratory Manual, 2nd edn. New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  27. Sattentau Q. J., Weiss R. A. 1988; The CD4 antigen: physiological ligand and HIV receptor. Cell 52:631–633
    [Google Scholar]
  28. Schawaller M., Smith G. E., Skehel J. J., Wiley D. C. 1989; Studies with crosslinking reagents on the oligomeric structure of the env glycoprotein of HIV. Virology 172:367–369
    [Google Scholar]
  29. Weiss C. D., Levy J. A., White J. M. 1990; Oligomeric organization of gp120 on infectious human immunodeficiency virus type I particles. Journal of Virology 64:5674–5677
    [Google Scholar]
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