Immunoglobulin E-binding site in Fc epsilon receptor (Fc epsilon RII/CD23) identified by homolog-scanning mutagenesis.
The IgE-binding site of the human low-affinity receptor for IgE (Fc epsilon RII/CD23) has previously been mapped to the extracellular domain between amino acid residues 160 and 287. We now have investigated which conformational epitope within this domain specifies the receptor-ligand interaction. The analysis of homolog-scanning mutants expressed in mammalian cells demonstrates that amino acid side chains that affect IgE binding are located in two discontinuous segments, between residues 165-190 and 224-256. The overall structure of the chimeric binding domains, as probed with 11 conformation-sensitive monoclonal antibodies, is generally not distorted, except by replacement of residues 165-183. In this region, disruption of binding function appears to be caused by global conformational constraints on the binding site. Substitution and deletion mutants demonstrate that six out of eight extracellular cysteines, Cys163, Cys174, Cys191, Cys259, Cys273, and Cys282, are necessary for IgE binding and are most likely involved in intramolecular disulfide bridges. We show that the Fc epsilon RII domain delineated by Cys163 and Cys282 encodes all the structural information required to form the IgE-binding site.
Bettler
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B
Department of Biotechnology, Ciba-Geigy Ltd., Basel, Switzerland.
Texido
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Raggini
S
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Rüegg
D
D
Hofstetter
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H
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D000944:Antigens, Differentiation, B-Lymphocyte / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites; D015153:Blotting, Western; D003545:Cysteine / Q000378:metabolism; D004220:Disulfides / Q000378:metabolism; D006801:Humans; D007073:Immunoglobulin E / Q000378:metabolism*; D008969:Molecular Sequence Data; D016296:Mutagenesis; D010641:Phenotype; D011487:Protein Conformation; D011961:Receptors, Fc / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D017455:Receptors, IgE; D012689:Sequence Homology, Nucleic Acid; D012694:Serine / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000944:Antigens, Differentiation, B-Lymphocyte; D004220:Disulfides; D011961:Receptors, Fc; D017455:Receptors, IgE; D007073:Immunoglobulin E; D012694:Serine; D003545:Cysteine