Chimeric Fc receptors identify immunoglobulin-binding regions in human Fc gamma RII and Fc epsilon RI.
Fc gamma RII and Fc epsilon RI are functionally distinct cell surface receptors for immunoglobulin (Ig); Fc gamma RII binds IgG with low affinity, whereas Fc epsilon RI binds IgE with high affinity, yet they are homologous in structure and sequence having extracellular regions containing two Ig-like domains with 38% amino acid identity. Chimeric receptors derived from human Fc gamma RII and Fc epsilon RI were produced by exchanging homologous regions of the two receptors to define binding region(s) for IgG in Fc gamma RII and IgE in Fc epsilon RI. Firstly, a chimeric form of the Fc epsilon RI alpha chain was produced by replacing the transmembrane region and cytoplasmic tail with that of Fc gamma RII. This mutant alpha chain could be expressed on the cell surface independently of associated beta and gamma subunits, and retained high-affinity IgE binding, indicating that the extracellular region of the Fc epsilon RI alpha chain is sufficient for high-affinity IgE binding. Secondly, to identify the role of the individual domains in Fc binding of both Fc gamma RII and Fc epsilon RI, chimeric receptors were generated by exchanging the first extracellular domains between Fc gamma RII and the alpha chain mutant and used to demonstrate that the second extracellular domain of both receptors contains region(s) directly involved in Ig binding. Additional chimeric receptors were constructed to localize the Ig interactive regions in domain two of Fc gamma RII and Fc epsilon RI; these identified a single region of IgG binding in Fc gamma RII located between residues Ser136 to Val169, and at least three independent IgE binding regions in the Fc epsilon RI alpha chain, between residues Trp87 to Lys128, Tyr129 to Asp145, and Ser146 to Val169.
Hulett
M D
MD
Austin Research Institute, Heidelberg, Australia.
McKenzie
I F
IF
Hogarth
P M
PM
D001483:Base Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D006801:Humans; D007073:Immunoglobulin E / Q000378:metabolism; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D017455:Receptors, IgE / Q000737:chemistry*; D017452:Receptors, IgG / Q000737:chemistry*; D011993:Recombinant Fusion Proteins; D012397:Rosette Formation
D009838:Oligodeoxyribonucleotides; D017455:Receptors, IgE; D017452:Receptors, IgG; D011993:Recombinant Fusion Proteins; D007073:Immunoglobulin E