The structural relation between the antigenic determinants to monoclonal antibodies and binding sites to rat brain synaptosomes and GT1b ganglioside in Clostridium botulinum type C neurotoxin.
The inhibition of the binding of 125I-labeled Clostridium botulinum type C neurotoxin to synaptosomes by unlabeled toxin indicated that there were two kinds of receptors on the synaptosomal membrane. The dissociation constants (Kd) were calculated as 79 pM and 35 nM from the concentration of unlabeled toxin that induced half-displacement of bound 125I-toxin. These values agree satisfactorily with the values obtained from direct binding experiments (Agui, T, Syuto, B., Oguma, K., Iida, H., & Kubo, S. (1983) J. Biochem. 94, 521-527). The inhibition of the binding of 125I-toxin to synaptosomes and N-acetylneuraminyl(alpha 2-3)galactosyl(beta 1-3)N-acetylgalactosaminyl(beta 1-4) [N-acetylneuraminyl(alpha 2-8) N-acetylneuraminyl(alpha 2-3)]galactosyl(beta 1-4)glucosyl(beta 1-1)ceramide (GT1b) by unlabeled heavy chain indicated that heavy chain facilitates the binding of toxin to synaptosomes and GT1b. The synaptosomal and heavy chain complex Kd values were estimated as 12 nM and 24 microM. Monoclonal antibodies C-9 and CA-12 recognized the binding sites to GT1b and synaptosomes, respectively. Antigenic determinants against the two antibodies are presumably partially overlapping, and the overlapping area seems to be essential to the reaction between toxin and C-9 antibody.
Agui
T
T
Syuto
B
B
Oguma
K
K
Iida
H
H
Kubo
S
S
D000818:Animals; D000911:Antibodies, Monoclonal / Q000502:physiology; D001921:Brain / Q000378:metabolism*; D000939:Epitopes / Q000276:immunology*; D005732:Gangliosides / Q000276:immunology / Q000378:metabolism*; D051381:Rats; D011950:Receptors, Cholinergic / Q000276:immunology / Q000378:metabolism; D013570:Synaptic Membranes / Q000378:metabolism*; D014121:Toxoids / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*
D000911:Antibodies, Monoclonal; C035415:Clostridium botulinum toxoid; D000939:Epitopes; D005732:Gangliosides; D011950:Receptors, Cholinergic; D014121:Toxoids; C016481:trisialoganglioside GT1