Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1986 Nov;83(21):8323–8327. doi: 10.1073/pnas.83.21.8323

Biological properties of a recombinant human immunoglobulin epsilon-chain fragment.

T Ishizaka, B Helm, J Hakimi, J Niebyl, K Ishizaka, H Gould
PMCID: PMC386920  PMID: 2430285

Abstract

A recombinant human immunoglobulin epsilon-chain gene expression product (rFc epsilon) was compared with a human E myeloma protein in the affinity for epsilon-chain Fc fragment receptors (Fc epsilon R) on cultured human basophils. The association-dissociation kinetics of the rFc epsilon-Fc epsilon R interaction are indistinguishable from that of E myeloma protein, indicating that rFc epsilon and IgE have identical affinity for the receptors. The recombinant gene product sensitizes cultured basophils for anti-IgE-induced histamine release. A dose-response curve of histamine release indicates that the gene product is equally efficient in transducing the signal for degranulation as the natural IgE. Both the rFc epsilon and IgE lost the affinity for Fc epsilon R by heating at 56 degrees C. Upon renaturation by passage through a solution of 6 M guanidine hydrochloride, rFc epsilon recovered both the affinity for Fc epsilon R and the original CD spectra. On the other hand, renaturation of heat-denatured IgE largely restored optical activity above 250 nm but restored neither the affinity for Fc epsilon R nor the CD spectrum below 220 nm. The results suggest that either the amino acid sequence or the carbohydrate present in the myeloma protein, but not the rFc epsilon, may interfere with refolding of the receptor-binding structures.

Full text

PDF
8323

Selected References

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

  1. 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]
  2. 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]
  3. 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]
  4. Ellsworth R. K., Nowak C. A. A gas chromatographic-mass spectrometric analysis of the esterifying alcohols of pumpkin seed protochlorophylls. Anal Biochem. 1974 Feb;57(2):534–546. doi: 10.1016/0003-2697(74)90108-0. [DOI] [PubMed] [Google Scholar]
  5. Geha R. S., Helm B., Gould H. Inhibition of the Prausnitz-Küstner reaction by an immunoglobulin epsilon-chain fragment synthesized in E. coli. Nature. 1985 Jun 13;315(6020):577–578. doi: 10.1038/315577a0. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Ishizaka K., Ishizaka T., Menzel A. E. Physicochemical properties of reaginic antibody. VI. Effect of heat on gamma-E-, gamma-G- and gamma-A-antibodies in the sera of ragweed sensitive patients. J Immunol. 1967 Sep;99(3):610–618. [PubMed] [Google Scholar]
  8. Ishizaka T., Conrad D. H., Huff T. F., Metcalfe D. D., Stevens R. L., Lewis R. A. Unique features of human basophilic granulocytes developed in in vitro culture. Int Arch Allergy Appl Immunol. 1985;77(1-2):137–143. doi: 10.1159/000233768. [DOI] [PubMed] [Google Scholar]
  9. Ishizaka T., Conrad D. H., Schulman E. S., Sterk A. R., Ishizaka K. Biochemical analysis of initial triggering events of IgE-mediated histamine release from human lung mast cells. J Immunol. 1983 May;130(5):2357–2362. [PubMed] [Google Scholar]
  10. Ishizaka T., Ishizaka K. Biology of immunoglobulin E. Molecular basis of reaginic hypersensitivity. Prog Allergy. 1975;19:60–121. [PubMed] [Google Scholar]
  11. Kenten J. H., Molgaard H. V., Houghton M., Derbyshire R. B., Viney J., Bell L. O., Gould H. J. Cloning and sequence determination of the gene for the human immunoglobulin epsilon chain expressed in a myeloma cell line. Proc Natl Acad Sci U S A. 1982 Nov;79(21):6661–6665. doi: 10.1073/pnas.79.21.6661. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Kulczycki A., Jr, Isersky C., Metzger H. The interaction of IgE with rat basophilic leukemia cells. I. Evidence for specific binding of IgE. J Exp Med. 1974 Mar 1;139(3):600–616. doi: 10.1084/jem.139.3.600. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kulczycki A., Jr, Metzger H. The interaction of IgE with rat basophilic leukemia cells. II. Quantitative aspects of the binding reaction. J Exp Med. 1974 Dec 1;140(6):1676–1695. doi: 10.1084/jem.140.6.1676. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kurokawa T., Seno M., Sasada R., Ono Y., Onda H., Igarashi K., Kikuchi M., Sugino Y., Honjo T. Expression of human immunoglobulin E epsilon chain cDNA in E. coli. Nucleic Acids Res. 1983 May 25;11(10):3077–3085. doi: 10.1093/nar/11.10.3077. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. McConahey P. J., Dixon F. J. A method of trace iodination of proteins for immunologic studies. Int Arch Allergy Appl Immunol. 1966;29(2):185–189. doi: 10.1159/000229699. [DOI] [PubMed] [Google Scholar]
  18. Ogawa M., Nakahata T., Leary A. G., Sterk A. R., Ishizaka K., Ishizaka T. Suspension culture of human mast cells/basophils from umbilical cord blood mononuclear cells. Proc Natl Acad Sci U S A. 1983 Jul;80(14):4494–4498. doi: 10.1073/pnas.80.14.4494. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES