Mutagenesis of immunoglobulin-like domains from the extracellular human interferon-gamma receptor alpha chain and their recognition by neutralizing antibodies monitored by surface plasmon resonance technology.
The extracellular portion of the human interferon-gamma receptor (IFN-gamma R) is predicted to adopt two Ig-like domains each with Ig superfamily type C2 topology. Surface plasmon resonance technology has been used here to compare the equilibrium and kinetic constants for binding reactions between these and several mutant domains, and neutralising anti-IFN-gamma R monoclonal antibodies. The biosensor measurements provide a sensitive method for monitoring the effects of mutations on the functional epitopes recognized by the neutralising antibodies. Thus, using recombinant native-like proteins made in E. coli, the ten N-terminal residues of the receptor were found not to be essential for domain folding, nor for recognition by the antibodies A6, D2 and gamma R38. In a similar way, residues in the interdomain region were found to play an important functional role in the epitope recognized by the antibody gamma R99.
Ruegg
N
N
Institute of Organic Chemistry, University of Zürich-Irchel, Switzerland.
Williams
G
G
Birch
A
A
Robinson
J A
JA
Schlatter
D
D
Huber
W
W
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology; D001483:Base Sequence; D015374:Biosensing Techniques; D007136:Immunoglobulins / Q000235:genetics* / Q000276:immunology*; D008565:Membrane Proteins / Q000032:analysis; D008969:Molecular Sequence Data; D016296:Mutagenesis / Q000235:genetics / Q000276:immunology; D016133:Polymerase Chain Reaction; D017471:Receptors, Interferon / Q000235:genetics / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology; D013057:Spectrum Analysis / Q000295:instrumentation / Q000379:methods*; D000096743:Interferon gamma Receptor
D000911:Antibodies, Monoclonal; D007136:Immunoglobulins; D008565:Membrane Proteins; D017471:Receptors, Interferon; D011994:Recombinant Proteins; D000096743:Interferon gamma Receptor