The translocation of botulinum A neurotoxin by chromaffin cells is promoted in low ionic strength solution and is insensitive to trypsin.
Botulinum A neurotoxin (BoNtx) produced a partial inhibition of carbachol induced 3H-noradrenaline (3H-NA) release from bovine adrenal chromaffin cells in monolayer culture. Each of the polysialogangliosides GD1a, GT1b and GD1b enhanced the block of exocytosis when they were applied prior to the toxin exposure. The monosialoganglioside GM1 was not effective. Chromaffin cells treated with neuraminidase lost their sensitivity to BoNtx. Application of gangliosides to these cells, however, restored their susceptibility to the toxin. Treatment of the cells with trypsin did not affect the BoNtx-blockade of 3H-NA-release. The potency of botulinum A toxin was increased in a solution of low ionic strength in which sodium chloride was replaced by sucrose. In agreement with the potency of botulinum A neurotoxin in blocking exocytosis under the various conditions, binding of 125I-botulinum A neurotoxin to chromaffin cells was enhanced in low ionic strength solution and by pretreatment of the cells with gangliosides. The binding was decreased by digestion of gangliosides with neuraminidase. It is concluded that botulinum A neurotoxin binds exclusively to polysialogangliosides which subsequently serve as carriers for the toxin. The low ionic strength may increase some physico-chemical interaction of the toxin with the polysialogangliosides.
Marxen
P
P
Medical School of Hannover, Dept of Pharmacology and Toxicology, F.R.G.
Erdmann
G
G
Bigalke
H
H
D000818:Animals; D001905:Botulinum Toxins / Q000378:metabolism*; D002478:Cells, Cultured; D002838:Chromaffin System / Q000166:cytology / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D005089:Exocytosis / Q000187:drug effects; D005732:Gangliosides / Q000494:pharmacology; D009439:Neuraminidase / Q000494:pharmacology; D009638:Norepinephrine / Q000378:metabolism; D012996:Solutions; D014357:Trypsin / Q000494:pharmacology*
D005732:Gangliosides; D012996:Solutions; D009439:Neuraminidase; D014357:Trypsin; D001905:Botulinum Toxins; D009638:Norepinephrine