Biochemistry (Moscow)

, Volume 74, Issue 2, pp 194–200 | Cite as

Dependence of conformation of D3/D4 domains of human CD4 on glycosylation and membrane attachment

  • L. F. LidemanEmail author
  • R. A. Gibadulin


Conformational dynamics of human T-helper cell receptor protein CD4 has been studied with the help of monoclonal antibody (mAb) T6. The mAb T6 discriminates between s- and m-forms of CD4 and recognizes a specific conformation of the soluble (s) form of CD4 including the first nine amino acids of CD4 transmembrane sequence. However, change of tryptophan for serine in position 2 in this sequence destabilizes the T6-type conformation. By enzymatic deglycosylation and deletions of glycosylation sites, we show that T6-type conformation depends on glycosylation in both sites (Asn271 and Asn300). We show also that the sugars are not involved in direct binding to the antibody but stabilize the D3/D4 local conformation. Deglycosylated forms of sCD4 in vivo acquire a specific conformation similar to the wild type sCD4, which however cannot be restored after denaturation/renaturation under conditions of non-reducing Western blot. This observation indicates that the correct protein folding needs chaperone assistance and cannot be achieved in vitro. Completely non-glycosylated sCD4 is synthesized and secreted into the growth medium. In the medium, this mutant appears to be unstable and aggregates during time. In a contrast to soluble CD4, mutations in glycosylation sites abrogate expression of membrane CD4, thus demonstrating a different secretion pathways for soluble and membrane proteins.

Key words

CD4 autoimmune epitope glycosylation protein folding HIV-1 



amino acid residue




monoclonal antibody


major histocompatibility complex


phosphate buffered saline

sCD4 and mCD4

soluble and membrane forms of CD4, respectively


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  1. 1.
    Abraham, N., Miceli, M. C., Parnes, J. R., and Veillette, A. (1991) Nature, 350, 62–66.PubMedCrossRefGoogle Scholar
  2. 2.
    Doyle, C., and Strominger, J. L. (1987) Nature, 330, 256–259.PubMedCrossRefGoogle Scholar
  3. 3.
    Reddy, M. M., Vodian, M., and Grieco, M. H. (1990) J. Clin. Microbiol., 28, 1744–1746.PubMedGoogle Scholar
  4. 4.
    Sweet, R. W., Truneh, A., and Hendrickson, W. A. (1991) Curr. Opin. Biotechnol., 2, 622–633.PubMedCrossRefGoogle Scholar
  5. 5.
    Carr, S. A., Hemling, M. E., Folena-Wasserman, G., Sweet, R. W., Anumula, K., Barr, J. R., Huddleston, M. J., and Taylor, P. (1989) J. Biol. Chem., 264, 21286–21295.PubMedGoogle Scholar
  6. 6.
    Peakman, M., Senaldi, G., Foote, N., McManus, T. J., and Vergani, D. (1992) J. Infect. Dis., 165, 799–804.PubMedGoogle Scholar
  7. 7.
    Yoneyama, A., Nakahara, K., Higashihara, M., and Kurokawa, K. (1995) Br. J. Haematol., 89, 47–54.PubMedGoogle Scholar
  8. 8.
    Symons, J. A., McCulloch, J. F., Wood, N. C., and Duff, G. W. (1991) Clin. Immunol. Immunopathol., 60, 72–82.PubMedCrossRefGoogle Scholar
  9. 9.
    Marcante, R., and Cavedon, G. (1991) Allergol. Immunopathol. (Madr.), 19, 99–102.Google Scholar
  10. 10.
    North, M. E., Spickett, G. P., Webster, A. D., and Farrant, J. (1991) Clin. Exp. Immunol., 86, 252–255.PubMedCrossRefGoogle Scholar
  11. 11.
    Matsumoto, Y., Shinzato, T., Takai, I., Nakai, S., Miwa, M., and Maeda, K. (1998) Nephron, 78, 490–491.PubMedCrossRefGoogle Scholar
  12. 12.
    Sato, S., Fujimoto, M., Kikuchi, K., Ihn, H., Tamaki, K., and Takehara, K. (1996) Arch. Dermatol. Res., 288, 358–362.PubMedCrossRefGoogle Scholar
  13. 13.
    Seagal, J., Spectorman, E., Gershoni, J. M., and Denisova, G. F. (2001) Transgenic Res., 10, 113–120.PubMedCrossRefGoogle Scholar
  14. 14.
    Denisova, G., Lideman, L., Spectorman, E., Abulafia-Lapid, R., Burke, M., Yust, I., and Gershoni, J. M. (2003) Mol. Immunol., 40, 231–239.PubMedCrossRefGoogle Scholar
  15. 15.
    Chams, V., Jouault, T., Fenouillet, E., Gluckman, J. C., and Klatzmann, D. (1988) Aids, 2, 353–361.PubMedCrossRefGoogle Scholar
  16. 16.
    Callahan, L. N., Roderiquez, G., Mallinson, M., and Norcross, M. A. (1992) J. Immunol., 149, 2194–2202.PubMedGoogle Scholar
  17. 17.
    Lideman, L. F., Kasennova, E. V., Denisova, G. F., Stakhanova, V. M., Zverev, S., Gibadulin, R. A., and Bobkov, A. F. (2005) Vopr. Virusol., 50, 15–19.PubMedGoogle Scholar
  18. 18.
    Gershoni, J. M., Denisova, G., Raviv, D., Smorodinsky, N. I., and Buyaner, D. (1993) FASEB J., 7, 1185–1187.PubMedGoogle Scholar
  19. 19.
    D’Aloja, P., Olivetta, E., Bona, R., Naooi, F., Pedacchia, D., Pugliese, K., Ferrari, G., Verani, P., and Federico, M. (1998) J. Virol., 72, 4308–4319.PubMedGoogle Scholar
  20. 20.
    Broder, C. C., and Berger, E. A. (1993) J. Virol., 67, 913–926.PubMedGoogle Scholar
  21. 21.
    Preusch, P. C., Norvell, J. C., Cassatt, J. C., and Cassman, M. (1998) Nat. Struct. Biol., 5, 12–14.PubMedCrossRefGoogle Scholar
  22. 22.
    Landolt-Marticorena, C., Williams, K. A., Deber, C. M., and Reithmeier, R. A. F. (1993) J. Mol. Biol., 229, 602–608.PubMedCrossRefGoogle Scholar
  23. 23.
    Reithmeier, R. A. F. (1995) Curr. Opin. Struct. Biol., 5, 491–500.PubMedCrossRefGoogle Scholar
  24. 24.
    De Planque, M. R., Kruijtzer, J. A., Liskamp, R. M., Marsh, D., Greathouse, D. V., Koeppe, R. E., de Kruijff II, B., and Killian, J. A. (1999) J. Biol. Chem., 274, 20839–20846.PubMedCrossRefGoogle Scholar
  25. 25.
    Yau, W. M., Wimley, W. C., Gawrisch, K., and White, S. H. (1988) Biochemistry, 37, 14713–14718.CrossRefGoogle Scholar
  26. 26.
    Ridder, A. N., Morein, S., Stam, J. G., Kuhn, A., de Kruijff, B., and Killian, J. A. (2000) Biochemistry, 39, 6521–6528.PubMedCrossRefGoogle Scholar
  27. 27.
    Manca, F., Seravalli, E., Valle, M. T., Fenoglio, D., Kunkl, A., Li Pira, G., Zolla-Pazner, S., and Celada, F. (1993) Int. Immunol., 5, 1109–1117.PubMedCrossRefGoogle Scholar
  28. 28.
    Schlueter, C., Hauke, S., Flohr, A. M., Rogalla, P., and Bullerdiek, J. (2003) Biochim. Biophys. Acta, 1630, 1–6.PubMedGoogle Scholar
  29. 29.
    Yang, J., and Liu, C. Q. (2000) Acta Pharmacol. Sin., 21, 547–553.PubMedGoogle Scholar
  30. 30.
    Mashikian, M. V., Tarpy, R. E., Saukkonen, J. J., Lim, K. G., Fine, G. D., Cruikshank, W. W., and Center, D. M. (1998) J. Allergy Clin. Immunol., 101, 786–792.PubMedCrossRefGoogle Scholar
  31. 31.
    Kaser, A., Dunzendorfer, S., Offner, F. A., Ryan, T., Schwabegger, A., Cruikshank, W. W., Wiedermann, C. J., and Tilg, H. (1999) J. Immunol., 163, 3232–3238.PubMedGoogle Scholar
  32. 32.
    Liu, Y., Cruikshank, W. W., O’Loughlin, T., O’Reilly, P., Center, D. M., and Kornfeld, H. (1999) J. Biol. Chem., 274, 23387–23395.PubMedCrossRefGoogle Scholar
  33. 33.
    Lynch, E. A., Heijens, C. A., Horst, N. F., Center, D. M., and Cruikshank, W. W. (2003) J. Immunol., 171, 4965–4968.PubMedGoogle Scholar
  34. 34.
    Watanabe, M., Iwatani, Y., Hidaka, Y., Mitsuda, N., and Amino, N. (1996) Am. J. Reprod. Immunol., 36, 220–227.PubMedGoogle Scholar

Copyright information

© Pleiades Publishing, Ltd. 2009

Authors and Affiliations

  1. 1.Ivanovsky Institute of VirologyRussian Academy of Medical SciencesMoscowRussia

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