Journal of Protein Chemistry

, Volume 12, Issue 4, pp 459–467 | Cite as

The disulfide bridges of the immunoglobulin-like domains of FcγRIIIB are essential for efficient expression and biological activity

  • Barbara Enenkel
  • Jürgen Frey


The immunoglobulin G receptor FcγRIIIB belongs to the immunoglobulin superfamily as two extracellular domains show homology to the immunoglobulin domains. Since some residues in these domains, such as the two cysteines, are supposed to form an intrachain disulfide bridge are so commonly conserved, they may be of importance for correct folding. Site-directed mutagenesis and expression in BHK21 confirmed this supposition for the FcγRIIIB. Replacing both cysteines in the first and/or second domain by serines reduced the surface expression level by 50%, whereas the ligand binding capability was 20–30% of that seen in cells expressing the wild-type receptor. Replacing one of the four cysteines resulted in the loss of surface expression. Exchanging the conserved tryptophan in the first domain by phenylalanine only slightly affected the ligand binding (25%), whereas the surface expression remained unchanged.

Key words

immunoglobulin G receptor CD16 conserved amino acids domain structure immunoglobulin-like domains intrachain disulfide bridges 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Allen, J.M., and Seed, B. (1988).Nucleic Acids Research 16, 11824.Google Scholar
  2. Allen, J.M., and Seed, B. (1989). Science243, 378–381.Google Scholar
  3. Anderson, C.L., and Looney, R.J. (1986).Immunology Today 7, 264–266.Google Scholar
  4. Anderson, C.L., Shen, L., Eicher, D.M., Wewers, M.D., and Gill, J.K. (1990).J. Exp. Med. 171, 1333–1345.Google Scholar
  5. Artelt, P., Morelle, C., Ausmeier, M., Fitzek, M., and Hauser, H. (1988). Gene68, 213–219.Google Scholar
  6. Ben-Sasson, S.Z., Le Gros, G., Conrad, D.H., Finkelman, F.D., and Paul, W.E. (1990).Proc. Natl. Acad. Sci. USA 87, 1421–1425.Google Scholar
  7. Brooks, D.G., Qiao Qiu W., Luster, A.D., and Ravetch J.V., (1989).J. Exp. Med. 170, 1369–1385.Google Scholar
  8. Burton, D.R., Jefferis, R., Partridge, L.J., and Woof, J.M. (1988).Moelcular Immunology 25, 1175–1181.Google Scholar
  9. Chen, E.Y., and Seeburg, P.H. (1985).DNA 4, 165–170.Google Scholar
  10. Chirgwin, J.M., Przybyla, A.E., MacDonald, R.J., and Rutter, W.J. (1979).Biochemistry 18, 5294–5299.Google Scholar
  11. Enenkel, B., Jung, D., and Frey, J. (1991).Eur. J. Immunol. 21, 659–663.Google Scholar
  12. Engelhardt, W., Geerds, C., and Frey, J. (1990).Eur. J. Immunol. 20, 1367–1377.Google Scholar
  13. Engelhardt, W., Gorczytza, H., Butterweck, A., Mönkemann, H., and Frey, J. (1991).Eur. J. Immunol. 21, 2227–2238.Google Scholar
  14. Fanger, M.W., Shen, L., Graziano, R.F., and Guyre, P.M. (1989).Immunology Today 10, 92–99.Google Scholar
  15. Fleit, H.B., Wright, S.D., and Unkeless, J.C. (1982).Proc. Natl. Acad. Sci. USA 79, 3275–3279.Google Scholar
  16. Frade, R., Barel, M., and Charriaut, C. (1983).Methods in Enzymology 93, 155–163.Google Scholar
  17. Frey, J., Janes, M., Englehardt, W., Afting, E.-G., Geerds, C., and Möller, B. (1986).Eur. J. Biochem. 158, 85–89.Google Scholar
  18. Graham, F.L., and Van der Eb, A.J. (1973).Virology 52, 456–467.Google Scholar
  19. Goto, Y., and Hamaguchi, K. (1979).J. Biochem. 86, 1433–1441.Google Scholar
  20. Haber, E. (1964).Proc. Natl. Acad. Sci. USA 52, 1099–1106.Google Scholar
  21. Halldén, G., Andersoon, H., Hed, J., and Johansson, S.G.O. (1989).J. Immunol. Meth. 124, 103–109.Google Scholar
  22. Heyman, B. (1990).Immunology Today 11, 310–313.Google Scholar
  23. Höning, S., Jockusch, B.M., Kreimer, G., Veltel, D., Robenek, H., Engelhardt, W., and Frey, J. (1991).Eur. J. Cell Biol. 55, 48–59.Google Scholar
  24. Huizinga, T.W.J., van der Schoot, C.E., Jost, C., Klaassen, R., Kleijer, M., von dem Borne, A.E.G.K., Roos, D., and Tetteroo, P.A.T. (1988).Nature 333, 667–669.Google Scholar
  25. Kinet, J.P. (1989).Cell 57, 351–354.Google Scholar
  26. Odin, J.A., Painter, C.J., and Unkeless, J.C. (1990).Cellular and Molecular Mechanisms of Inflammation. (Cochrane, C.G., and Gimbrone, M.A., eds.), Vol. 1, pp. 1–33.Google Scholar
  27. Odin, J.A., Edberg, J.C., Painter, C.J., Kimberley, R.P., and Unkeless, J.C. (1991).Science 254, 1785–1788.Google Scholar
  28. Ravetch, J.V., and Perussia, B. (1989).J. Exp. Med. 170, 481–497.Google Scholar
  29. Ravetch, J.V., and Kinet, J.P. (1991).Ann. Rev. Immunol. 9, 457–492.Google Scholar
  30. Rigley, K.P., Harnett, M.M., and Klaus, G.G.B. (1989).Eur. J. Immunol. 19, 481–485.Google Scholar
  31. Rudikoff, S., and Pumphrey, J.G (1986).Proc. Natl. Acad. Sci. USA 83, 7875–7878.Google Scholar
  32. Saiki, R.K., Gelfand, D.H., Stoffel, S., Scharf, S.J., Higuchi, R., Horn, G.T., Mullis, K.B., and Erlich, H.A. (1988).Science 239, 487–491.Google Scholar
  33. Sambrook, J., Fritsch, E.F., and Maniatis, T. (1989).Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press.Google Scholar
  34. Sanger, F., Nicklen, S., and Coulson, A.R. (1977).Proc. Natl. Acad. Sci. USA 74, 5463–5467.Google Scholar
  35. Sautès, C., Varin, N., Hogarth, P.M., Unkeless, J.C., Teillaud, C., Even, J., Lynch, A., and Fridman, W.H. (1990).Molecular Immunology 27, 1201–1207.Google Scholar
  36. Scallon, B.J., Scigliano, E., Freeman, V.H., Miedel, M.C., Pan, Y.-C.E., Unkeless, J.C., and Kochan, J.P. (1989).Proc. Natl. Acad. Sci. USA 86, 5079–5083.Google Scholar
  37. Selvaraj, P., Rosse, W.F., Silber, R., and Springer, T.A. (1988).Nature 333, 565–567.Google Scholar
  38. Shiroishi, T., Evans, G.A., Appella, E., and Ozato, K. (1984).Proc. Natl. Acad. Sci. USA 81, 7544–7548.Google Scholar
  39. Simmons, D., and Seed, B. (1988).Nature 333, 568–570.Google Scholar
  40. Sinclair, N.R.S., and Panoskaltsis, A. (1987).Immunology Today 8, 76–79.Google Scholar
  41. Stanssens, P., Opsomer, C., McKeown, Y.M., Kramer, W., Zabeau, M., and Fritz, H.-J. (1989).Nucleic Acids Research 17, 4441–4454.Google Scholar
  42. Stengelin, S., Stamenkovic, I., and Seed, B. (1988).EMBO J. 8, 1053–1059.Google Scholar
  43. Stuart, S.G., Simister, N.E., Clarkson, S.B., Kacinski, B.M., Shapiro, M., and Mellman, I. (1989).EMBO J. 8, 3657–3666.Google Scholar
  44. Vara, J.A., Portela, A., Ortin, J., and Jiménez, A. (1986).Nucleic Acids Research 14, 4617–4624.Google Scholar
  45. Varin, N., Sautès, C., Galinha, A., Even, J., Hogarth, P.M., and Fridman, W.H. (1989).Eur. J. Immunol. 19, 2263–2268.Google Scholar
  46. Vieira, J., and Messing, J. (1987).Methods in Enzymology 153, 3–11.Google Scholar
  47. Williams, A.F. (1987).Immunology Today 8, 298–303.Google Scholar
  48. Williams, A.F., and Barclay, A.N. (1988).Ann. Rev. Immunol. 6, 381–405.Google Scholar

Copyright information

© Plenum Publishing Corporation 1993

Authors and Affiliations

  • Barbara Enenkel
    • 1
  • Jürgen Frey
    • 1
  1. 1.Fakultät für Chemie, Biochemie IIUniversität BielefeldBielefeld 1Germany

Personalised recommendations