Processing of tetanus and botulinum A neurotoxins in isolated chromaffin cells.
Tetanus and botulinum A neurotoxins were introduced into the cytosol of chromaffin cells by means of an electric field in which the plasma membrane is forced to form pores of approximately 1 micron at the sites facing the electrodes. As demonstrated by electron microscopy, both [125I] and gold-labelled tetanus toxin (TeTx) diffuse through these transient openings. Dichain-TeTx, with its light chain linked to the heavy chain by means of a disulfide bond, causes the block of exocytosis to develop more slowly than does the purified light chain. The disulfide bonds, which in both toxins hold the subunits together, were cleaved by the intrinsic thioredoxin-reductase system. Single chain TeTx, in which the heavy and light chains are interconnected by an additional peptide bond, was far less effective than dichain-TeTx at blocking exocytosis, which indicates that proteolysis is the rate-limiting step. The toxins were degraded further to low-molecular weight fragments which, together with intact toxins and subunits, were released by the cells. The intracellular half-life of [125I] dichain-TeTx was approximately three days. The number of light-chain molecules required to maintain exocytosis block in a single cell, as calculated by two different methods, was less than 10. The long duration of tetanus poisoning may result from the persistence of intracellular toxin due to scarcity of free cytosolic proteases. This may also hold for the slow recovery from botulism.
Erdal
E
E
PharmaTec GmbH, Flörsheim, Germany.
Bartels
F
F
Binscheck
T
T
Erdmann
G
G
Frevert
J
J
Kistner
A
A
Weller
U
U
Wever
J
J
Bigalke
H
H
D000313:Adrenal Medulla / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D000818:Animals; D001905:Botulinum Toxins / Q000378:metabolism* / Q000494:pharmacology; D002417:Cattle; D002463:Cell Membrane Permeability; D002478:Cells, Cultured; D018274:Electroporation; D005089:Exocytosis / Q000502:physiology; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D013744:Tetanus Toxin / Q000378:metabolism* / Q000494:pharmacology
D013744:Tetanus Toxin; D001905:Botulinum Toxins