Mapping of the high affinity Fc epsilon receptor binding site to the third constant region domain of IgE.
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.
Nissim
A
A
Department of Chemical Immunology, Weizmann Institute of Science, Rehovot, Israel.
Jouvin
M H
MH
Eshhar
Z
Z
D000818:Animals; D000944:Antigens, Differentiation, B-Lymphocyte / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites; D001667:Binding, Competitive; D002872:Chromosome Deletion; D005091:Exons; D006801:Humans; D007127:Immunoglobulin Constant Regions / Q000235:genetics / Q000378:metabolism*; D007073:Immunoglobulin E / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D051379:Mice; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D010957:Plasmids; D011961:Receptors, Fc / Q000378:metabolism*; D017455:Receptors, IgE; D011994:Recombinant Proteins / Q000378:metabolism; D015183:Restriction Mapping
D000944:Antigens, Differentiation, B-Lymphocyte; D007127:Immunoglobulin Constant Regions; D015345:Oligonucleotide Probes; D011961:Receptors, Fc; D017455:Receptors, IgE; D011994:Recombinant Proteins; D007073:Immunoglobulin E