Binding of Clostridium botulinum type C neurotoxin to different neuroblastoma cell lines.
Binding of type C neurotoxin (C1 toxin) from Clostridium botulinum (strain Stockholm) to neuroblastoma cell lines was studied by using biotinylated anti-toxin antibody and avidin-biotinylated peroxidase complex. The neurotoxin bound with high efficiency to mouse neuroblastoma (NS-20Y and NIE-115) cells and to hybridomas of rat glioblastoma and mouse neuroblastoma (NG108-C15) cells. The toxin bound little to human neuroblastoma, rat astrocytoma, and nonneural cell lines. Binding of the neurotoxin to NG108-C15 cells was inhibited by gangliosides (GT1b and GM1) and by monoclonal antibodies (CA-12 and C-9), although inhibition was not complete. Sequential preincubation of C1 toxin with GT1b and CA-12 caused complete inhibition. A Scatchard plot of binding of 125I-labeled C1 toxin to NG108-C15 cells showed a hyperbolic curve. Monoclonal antibody CA-12 but not C-9 neutralized the lethal activity of the toxin toward mice. Only C-9 clearly inhibited toxin binding to GT1b. These results suggest that NG108-C15 cells have at least two kinds of receptors for C1 toxin. From the results of binding tests with neuraminidase-, pronase-, and trypsin-treated NG108-C15 cells, the chemical nature of the high-affinity site was presumed to be a glycoprotein containing sialic acid. GT1b may have an important role in low-affinity sites.
Yokosawa
N
N
Department of Microbiology, Sapporo Medical College, Japan.
Kurokawa
Y
Y
Tsuzuki
K
K
Syuto
B
B
Fujii
N
N
Kimura
K
K
Oguma
K
K
D000818:Animals; D000911:Antibodies, Monoclonal; D001667:Binding, Competitive; D001905:Botulinum Toxins / Q000276:immunology / Q000378:metabolism*; D002460:Cell Line; D005732:Gangliosides / Q000494:pharmacology; D005910:Glioma / Q000378:metabolism; D006801:Humans; D006822:Hybrid Cells / Q000378:metabolism; D007136:Immunoglobulins; D007700:Kinetics; D051379:Mice; D009447:Neuroblastoma / Q000378:metabolism*; D051381:Rats; D011950:Receptors, Cholinergic / Q000032:analysis* / Q000187:drug effects / Q000276:immunology
D000911:Antibodies, Monoclonal; D005732:Gangliosides; D007136:Immunoglobulins; D011950:Receptors, Cholinergic; D001905:Botulinum Toxins