Affinity purified tetanus toxin binds to isolated chromaffin granules and inhibits catecholamine release in digitonin-permeabilized chromaffin cells.
Tetanus toxin, a potent neurotoxin which blocks neurotransmitter release in the CNS, also inhibits Ca2+-induced catecholamine release from digitonin-permeabilized, but not from intact bovine chromaffin cells. In searching for intracellular targets for the toxin we studied the binding of affinity-purified tetanus toxin to bovine adrenal chromaffin granules. Tetanus toxin bound in a neuraminidase-sensitive fashion to intact granules and to isolated granule membranes, as assayed biochemically and visualized by electron microscopic techniques. The binding characteristics of the toxin to chromaffin granule membranes are very similar to the binding of tetanus toxin to brain synaptosomal membranes. We suggest that the toxin-binding site is a glycoconjugate of the G1b type (a polysialoganglioside or a glycoprotein-proteoglycan) which is localized on the cytoplasmic face of the granule membrane and might directly be involved in exocytotic membrane fusion.
Lazarovici
P
P
Department of Pharmacology and Experimental Therapeutics, Hadassah School of Medicine, Jerusalem, Israel.
Fujita
K
K
Contreras
M L
ML
DiOrio
J P
JP
Lelkes
P I
PI
D000313:Adrenal Medulla / Q000378:metabolism*; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002395:Catecholamines / Q000378:metabolism*; D002417:Cattle; D002463:Cell Membrane Permeability / Q000187:drug effects; D002837:Chromaffin Granules / Q000378:metabolism*; D002838:Chromaffin System / Q000378:metabolism*; D004072:Digitonin / Q000494:pharmacology; D006001:Glycoconjugates / Q000378:metabolism; D066298:In Vitro Techniques; D008854:Microscopy, Electron; D009439:Neuraminidase / Q000494:pharmacology; D012636:Secretory Rate / Q000187:drug effects; D013744:Tetanus Toxin / Q000378:metabolism*
D002395:Catecholamines; D006001:Glycoconjugates; D013744:Tetanus Toxin; D009439:Neuraminidase; D004072:Digitonin; D002118:Calcium