Abstract
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.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Allen, J.M., and Seed, B. (1988).Nucleic Acids Research 16, 11824.
Allen, J.M., and Seed, B. (1989). Science243, 378–381.
Anderson, C.L., and Looney, R.J. (1986).Immunology Today 7, 264–266.
Anderson, C.L., Shen, L., Eicher, D.M., Wewers, M.D., and Gill, J.K. (1990).J. Exp. Med. 171, 1333–1345.
Artelt, P., Morelle, C., Ausmeier, M., Fitzek, M., and Hauser, H. (1988). Gene68, 213–219.
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.
Brooks, D.G., Qiao Qiu W., Luster, A.D., and Ravetch J.V., (1989).J. Exp. Med. 170, 1369–1385.
Burton, D.R., Jefferis, R., Partridge, L.J., and Woof, J.M. (1988).Moelcular Immunology 25, 1175–1181.
Chen, E.Y., and Seeburg, P.H. (1985).DNA 4, 165–170.
Chirgwin, J.M., Przybyla, A.E., MacDonald, R.J., and Rutter, W.J. (1979).Biochemistry 18, 5294–5299.
Enenkel, B., Jung, D., and Frey, J. (1991).Eur. J. Immunol. 21, 659–663.
Engelhardt, W., Geerds, C., and Frey, J. (1990).Eur. J. Immunol. 20, 1367–1377.
Engelhardt, W., Gorczytza, H., Butterweck, A., Mönkemann, H., and Frey, J. (1991).Eur. J. Immunol. 21, 2227–2238.
Fanger, M.W., Shen, L., Graziano, R.F., and Guyre, P.M. (1989).Immunology Today 10, 92–99.
Fleit, H.B., Wright, S.D., and Unkeless, J.C. (1982).Proc. Natl. Acad. Sci. USA 79, 3275–3279.
Frade, R., Barel, M., and Charriaut, C. (1983).Methods in Enzymology 93, 155–163.
Frey, J., Janes, M., Englehardt, W., Afting, E.-G., Geerds, C., and Möller, B. (1986).Eur. J. Biochem. 158, 85–89.
Graham, F.L., and Van der Eb, A.J. (1973).Virology 52, 456–467.
Goto, Y., and Hamaguchi, K. (1979).J. Biochem. 86, 1433–1441.
Haber, E. (1964).Proc. Natl. Acad. Sci. USA 52, 1099–1106.
Halldén, G., Andersoon, H., Hed, J., and Johansson, S.G.O. (1989).J. Immunol. Meth. 124, 103–109.
Heyman, B. (1990).Immunology Today 11, 310–313.
Höning, S., Jockusch, B.M., Kreimer, G., Veltel, D., Robenek, H., Engelhardt, W., and Frey, J. (1991).Eur. J. Cell Biol. 55, 48–59.
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.
Kinet, J.P. (1989).Cell 57, 351–354.
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.
Odin, J.A., Edberg, J.C., Painter, C.J., Kimberley, R.P., and Unkeless, J.C. (1991).Science 254, 1785–1788.
Ravetch, J.V., and Perussia, B. (1989).J. Exp. Med. 170, 481–497.
Ravetch, J.V., and Kinet, J.P. (1991).Ann. Rev. Immunol. 9, 457–492.
Rigley, K.P., Harnett, M.M., and Klaus, G.G.B. (1989).Eur. J. Immunol. 19, 481–485.
Rudikoff, S., and Pumphrey, J.G (1986).Proc. Natl. Acad. Sci. USA 83, 7875–7878.
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.
Sambrook, J., Fritsch, E.F., and Maniatis, T. (1989).Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press.
Sanger, F., Nicklen, S., and Coulson, A.R. (1977).Proc. Natl. Acad. Sci. USA 74, 5463–5467.
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.
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.
Selvaraj, P., Rosse, W.F., Silber, R., and Springer, T.A. (1988).Nature 333, 565–567.
Shiroishi, T., Evans, G.A., Appella, E., and Ozato, K. (1984).Proc. Natl. Acad. Sci. USA 81, 7544–7548.
Simmons, D., and Seed, B. (1988).Nature 333, 568–570.
Sinclair, N.R.S., and Panoskaltsis, A. (1987).Immunology Today 8, 76–79.
Stanssens, P., Opsomer, C., McKeown, Y.M., Kramer, W., Zabeau, M., and Fritz, H.-J. (1989).Nucleic Acids Research 17, 4441–4454.
Stengelin, S., Stamenkovic, I., and Seed, B. (1988).EMBO J. 8, 1053–1059.
Stuart, S.G., Simister, N.E., Clarkson, S.B., Kacinski, B.M., Shapiro, M., and Mellman, I. (1989).EMBO J. 8, 3657–3666.
Vara, J.A., Portela, A., Ortin, J., and Jiménez, A. (1986).Nucleic Acids Research 14, 4617–4624.
Varin, N., Sautès, C., Galinha, A., Even, J., Hogarth, P.M., and Fridman, W.H. (1989).Eur. J. Immunol. 19, 2263–2268.
Vieira, J., and Messing, J. (1987).Methods in Enzymology 153, 3–11.
Williams, A.F. (1987).Immunology Today 8, 298–303.
Williams, A.F., and Barclay, A.N. (1988).Ann. Rev. Immunol. 6, 381–405.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Enenkel, B., Frey, J. The disulfide bridges of the immunoglobulin-like domains of FcγRIIIB are essential for efficient expression and biological activity. J Protein Chem 12, 459–467 (1993). https://doi.org/10.1007/BF01025046
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF01025046