Immunological neutralization of cobrotoxin by its homologous precipitin and non-precipitin antibodies.
Anti-cobrotoxin antibodies can be separated into precipitin and non-precipitin antibodies. The precipitin antibody possesses the same binding affinity to cobrotoxin as non-precipitin antibody, but the neutralizing capability of the latter is superior to that of the former in blocking cobrotoxin binding to nicotinic acetylcholine receptor (nAChR). After preincubation with antibodies, cobrotoxin completely lost its binding activity to nAChR. The dissociation of cobrotoxin-nAChR complex by the antibodies was low, and 60% of the complex formation appeared to be irreversible. These results indicate that the neutralization of cobrotoxin by the antibody may predominantly involve unbound, receptor-free cobrotoxin. The relationships of neutralization capacity and binding affinity as well as bond strength between cobrotoxin and its antibodies are incongruous. Different local conformational changes of a unique Trp in cobrotoxin on binding with the precipitin and non-precipitin antibodies seem to lead to different accessibility for fluorescence quenchers. Characterization of the binding domains by immunoprecipitation with the antibodies correlated with the quenching results. Thus, the binding topography of cobrotoxin may play an important role over the binding affinity and bond strength in neutralization by cobrotoxin antibody.
Kuo
K W
KW
Department of Biochemistry, Kaohsiung Medical College, Taiwan.
Chang
L S
LS
Lin
P M
PM
Leaber
R J
RJ
Chang
C C
CC
D000818:Animals; D000906:Antibodies / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology*; D000915:Antibody Affinity; D001666:Binding Sites, Antibody; D003039:Cobra Neurotoxin Proteins / Q000276:immunology* / Q000378:metabolism; D007700:Kinetics; D009500:Neutralization Tests; D011234:Precipitins / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology*; D011487:Protein Conformation; D011978:Receptors, Nicotinic / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D014101:Torpedo; D014364:Tryptophan / Q000032:analysis
D000906:Antibodies; D003039:Cobra Neurotoxin Proteins; D011234:Precipitins; D011978:Receptors, Nicotinic; D014364:Tryptophan