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. 1991 Jan;10(1):101–107. doi: 10.1002/j.1460-2075.1991.tb07925.x

Mapping of the high affinity Fc epsilon receptor binding site to the third constant region domain of IgE.

A Nissim 1, M H Jouvin 1, Z Eshhar 1
PMCID: PMC452617  PMID: 1824934

Abstract

Identification of the precise region(s) on the IgE molecule that take part in the binding of IgE to its high affinity receptor (Fc epsilon RI) may lead to the design of IgE analogues able to block the allergic response. To localize the Fc epsilon RI-binding domain of mouse IgE, we attempted to confer on human IgE, which normally does not bind to the rodent receptor, the ability to bind to the rat Fc epsilon RI. Employing exon shuffling, we have expressed chimeric epsilon-heavy chain genes composed of a mouse (4-hydroxy-3-nitrophenyl)acetic acid (NP)-binding VH domain, and human C epsilon in which various domains were replaced by their murine counterparts. This has enabled us to test the Fc epsilon RI-binding of each mouse IgE domain while maintaining the overall conformation of the molecule. All of the chimeric IgE molecules which contain the murine C epsilon 3, bound equally to both the rodent and human receptor, as well as to monoclonal antibodies recognizing a site on IgE which is identical or very close to the Fc epsilon RI binding site. Deletion of the second constant region domain did not impair either the binding capacity of the mutated IgE or its ability to mediate mast cell degradation. These results assign the third epsilon domain of IgE as the principal region involved in the interaction with the Fc epsilon RI.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Baird B., Holowka D. Structural mapping of Fc receptor bound immunoglobulin E: proximity to the membrane surface of the antibody combining site and another site in the Fab segments. Biochemistry. 1985 Oct 22;24(22):6252–6259. doi: 10.1021/bi00343a032. [DOI] [PubMed] [Google Scholar]
  2. Baniyash M., Eshhar Z., Rivnay B. Relationships between epitopes on IgE recognized by defined monoclonal antibodies and by the FC epsilon receptor on basophils. J Immunol. 1986 Jan;136(2):588–593. [PubMed] [Google Scholar]
  3. Baniyash M., Kehry M., Eshhar Z. Anti-IgE monoclonal antibodies directed at the Fc epsilon receptor binding site. Mol Immunol. 1988 Aug;25(8):705–711. doi: 10.1016/0161-5890(88)90106-x. [DOI] [PubMed] [Google Scholar]
  4. Barsumian E. L., Isersky C., Petrino M. G., Siraganian R. P. IgE-induced histamine release from rat basophilic leukemia cell lines: isolation of releasing and nonreleasing clones. Eur J Immunol. 1981 Apr;11(4):317–323. doi: 10.1002/eji.1830110410. [DOI] [PubMed] [Google Scholar]
  5. Bennich H., Ragnarsson U., Johansson S. G., Ishizaka K., Ishizaka T., Levy D. A., Lichtenstein L. M. Failure of the putative IgE pentapeptide to compete with IgE for receptors on basophils and masts cells. Int Arch Allergy Appl Immunol. 1977;53(5):459–468. doi: 10.1159/000231784. [DOI] [PubMed] [Google Scholar]
  6. Blank U., Ra C., Miller L., White K., Metzger H., Kinet J. P. Complete structure and expression in transfected cells of high affinity IgE receptor. Nature. 1989 Jan 12;337(6203):187–189. doi: 10.1038/337187a0. [DOI] [PubMed] [Google Scholar]
  7. Burt D. S., Hastings G. Z., Healy J., Stanworth D. R. Analysis of the interaction between rat immunoglobulin E and rat mast cells using anti-peptide antibodies. Mol Immunol. 1987 Apr;24(4):379–389. doi: 10.1016/0161-5890(87)90180-5. [DOI] [PubMed] [Google Scholar]
  8. Burt D. S., Stanworth D. R. Inhibition of binding of rat IgE to rat mast cells by synthetic IgE peptides. Eur J Immunol. 1987 Mar;17(3):437–440. doi: 10.1002/eji.1830170323. [DOI] [PubMed] [Google Scholar]
  9. Coleman J. W., Helm B. A., Stanworth D. R., Gould H. J. Inhibition of mast cell sensitization in vitro by a human immunoglobulin epsilon-chain fragment synthesized in Escherichia coli. Eur J Immunol. 1985 Sep;15(9):966–969. doi: 10.1002/eji.1830150920. [DOI] [PubMed] [Google Scholar]
  10. Conrad D. H., Wingard J. R., Ishizaka T. The interaction of human and rodent IgE with the human basophil IgE receptor. J Immunol. 1983 Jan;130(1):327–333. [PubMed] [Google Scholar]
  11. Dorrington K. J., Bennich H. H. Structure-function relationships in human immunoglobulin E. Immunol Rev. 1978;41:3–25. doi: 10.1111/j.1600-065x.1978.tb01458.x. [DOI] [PubMed] [Google Scholar]
  12. Dorrington K. J., Bennich H. Thermally induced structural changes in immunoglobulin E. J Biol Chem. 1973 Dec 25;248(24):8378–8384. [PubMed] [Google Scholar]
  13. Eshhar Z., Ofarim M., Waks T. Generation of hybridomas secreting murine reaginic antibodies of anti-DNP specificity. J Immunol. 1980 Feb;124(2):775–780. [PubMed] [Google Scholar]
  14. Flanagan J. G., Rabbitts T. H. The sequence of a human immunoglobulin epsilon heavy chain constant region gene, and evidence for three non-allelic genes. EMBO J. 1982;1(5):655–660. doi: 10.1002/j.1460-2075.1982.tb01223.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hamburger R. N. Peptide inhibition of the Prausnitz-Küstner reaction. Science. 1975 Aug 1;189(4200):389–390. doi: 10.1126/science.1145208. [DOI] [PubMed] [Google Scholar]
  16. Helm B., Kebo D., Vercelli D., Glovsky M. M., Gould H., Ishizaka K., Geha R., Ishizaka T. Blocking of passive sensitization of human mast cells and basophil granulocytes with IgE antibodies by a recombinant human epsilon-chain fragment of 76 amino acids. Proc Natl Acad Sci U S A. 1989 Dec;86(23):9465–9469. doi: 10.1073/pnas.86.23.9465. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Helm B., Marsh P., Vercelli D., Padlan E., Gould H., Geha R. The mast cell binding site on human immunoglobulin E. Nature. 1988 Jan 14;331(6152):180–183. doi: 10.1038/331180a0. [DOI] [PubMed] [Google Scholar]
  18. Ishida N., Ueda S., Hayashida H., Miyata T., Honjo T. The nucleotide sequence of the mouse immunoglobulin epsilon gene: comparison with the human epsilon gene sequence. EMBO J. 1982;1(9):1117–1123. doi: 10.1002/j.1460-2075.1982.tb01306.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Ishizaka K., Ishizaka T., Lee E. H. Biologic function of the Fc fragments of E myeloma protein. Immunochemistry. 1970 Aug;7(8):687–702. doi: 10.1016/0019-2791(70)90175-8. [DOI] [PubMed] [Google Scholar]
  20. Ishizaka T., Ishizaka K. Biology of immunoglobulin E. Molecular basis of reaginic hypersensitivity. Prog Allergy. 1975;19:60–121. [PubMed] [Google Scholar]
  21. Kenten J., Helm B., Ishizaka T., Cattini P., Gould H. Properties of a human immunoglobulin epsilon-chain fragment synthesized in Escherichia coli. Proc Natl Acad Sci U S A. 1984 May;81(10):2955–2959. doi: 10.1073/pnas.81.10.2955. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Küster H., Thompson H., Kinet J. P. Characterization and expression of the gene for the human Fc receptor gamma subunit. Definition of a new gene family. J Biol Chem. 1990 Apr 15;265(11):6448–6452. [PubMed] [Google Scholar]
  23. Liu F. T., Albrandt K. A., Bry C. G., Ishizaka T. Expression of a biologically active fragment of human IgE epsilon chain in Escherichia coli. Proc Natl Acad Sci U S A. 1984 Sep;81(17):5369–5373. doi: 10.1073/pnas.81.17.5369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Liu F. T., Albrandt K., Sutcliffe J. G., Katz D. H. Cloning and nucleotide sequence of mouse immunoglobulin epsilon chain cDNA. Proc Natl Acad Sci U S A. 1982 Dec;79(24):7852–7856. doi: 10.1073/pnas.79.24.7852. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Neuberger M. S., Williams G. T., Mitchell E. B., Jouhal S. S., Flanagan J. G., Rabbitts T. H. A hapten-specific chimaeric IgE antibody with human physiological effector function. Nature. 1985 Mar 21;314(6008):268–270. doi: 10.1038/314268a0. [DOI] [PubMed] [Google Scholar]
  26. Nishida Y., Miki T., Hisajima H., Honjo T. Cloning of human immunoglobulin epsilon chain genes: evidence for multiple C epsilon genes. Proc Natl Acad Sci U S A. 1982 Jun;79(12):3833–3837. doi: 10.1073/pnas.79.12.3833. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Oi V. T., Morrison S. L., Herzenberg L. A., Berg P. Immunoglobulin gene expression in transformed lymphoid cells. Proc Natl Acad Sci U S A. 1983 Feb;80(3):825–829. doi: 10.1073/pnas.80.3.825. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Perez-Montfort R., Metzger H. Proteolysis of soluble IgE-receptor complexes: localization of sites on IgE which interact with the Fc receptor. Mol Immunol. 1982 Sep;19(9):1113–1125. doi: 10.1016/0161-5890(82)90322-4. [DOI] [PubMed] [Google Scholar]
  29. Potter H., Weir L., Leder P. Enhancer-dependent expression of human kappa immunoglobulin genes introduced into mouse pre-B lymphocytes by electroporation. Proc Natl Acad Sci U S A. 1984 Nov;81(22):7161–7165. doi: 10.1073/pnas.81.22.7161. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rittenberg M. B., Pratt K. L. Antitrinitrophenyl (TNP) plaque assay. Primary response of Balb/c mice to soluble and particulate immunogen. Proc Soc Exp Biol Med. 1969 Nov;132(2):575–581. doi: 10.3181/00379727-132-34264. [DOI] [PubMed] [Google Scholar]
  31. Robertson M. W., Liu F. T. IgE structure-function relationships defined by sequence directed antibodies induced by synthetic peptides. Mol Immunol. 1988 Feb;25(2):103–113. doi: 10.1016/0161-5890(88)90057-0. [DOI] [PubMed] [Google Scholar]
  32. Rousseaux-Prévost R., Rousseaux J., Bazin H., Biserte G. Differential reduction of the inter-chain disulfide bonds of rat immunoglobulin E: relation to biological activity. Mol Immunol. 1984 Mar;21(3):233–241. doi: 10.1016/0161-5890(84)90078-6. [DOI] [PubMed] [Google Scholar]
  33. Schwarzbaum S., Nissim A., Alkalay I., Ghozi M. C., Schindler D. G., Bergman Y., Eshhar Z. Mapping of murine IgE epitopes involved in IgE-Fc epsilon receptor interactions. Eur J Immunol. 1989 Jun;19(6):1015–1023. doi: 10.1002/eji.1830190610. [DOI] [PubMed] [Google Scholar]
  34. Stanworth D. R., Humphrey J. H., Bennich H., Johansson S. G. Inhibition of Prausnitz-Küstner reaction by proteolytic-cleavage fragments of a human myeloma protein of immunoglobulin class E. Lancet. 1968 Jul 6;2(7558):17–18. doi: 10.1016/s0140-6736(68)92889-4. [DOI] [PubMed] [Google Scholar]

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