Inhibition of vacuolar adenosine triphosphatase antagonizes the effects of clostridial neurotoxins but not phospholipase A2 neurotoxins.
Bafilomycin A1, an inhibitor of vacuolar adenosine triphosphatase, was tested for its ability to antagonize botulinum neurotoxins (serotypes A-G), tetanus toxin and phospholipase A2 neurotoxins (notexin, beta-bungarotoxin, taipoxin and textilotoxin) on the mouse phrenic nerve-hemidiaphragm preparation. Bafilomycin itself produced concentration-dependent blockade of neuromuscular transmission without blocking nerve action potentials or muscle action potentials. This effect may have been due to inhibition of the proton pump that regulates acetylcholine transport into vesicles. At submaximal concentrations, bafilomycin was very effective in delaying the onset of paralysis due to all clostridial neurotoxins, but it had no protective effect against phospholipase A2 neurotoxins. Experiments were done to determine which of the three steps in clostridial neurotoxin action was antagonized by bafilomycin (e.g., binding, internalization and intracellular poisoning). Both pharmacological experiments and ligand-binding experiments showed that the drug did not block toxin binding to the plasma membrane. Similarly, pharmacological experiments on the time-dependent effects of bafilomycin showed that the drug did not antagonize the intracellular actions of toxins. The data indicated that bafilomycin acted at the intermediate step of internalization. This is in keeping with the facts that: 1) bafilomycin inhibits vacuolar adenosine triphosphatase, which in turn leads to inhibition of acidification in endosomes and 2) clostridial neurotoxins depend upon acidification of endosomes for translocation to the cytosol. The finding that bafilomycin antagonizes tetanus toxin may provide important clues for understanding how this toxin can act locally to produce flaccid paralysis. The finding that bafilomycin is a universal antagonist that protects against all clostridial neurotoxins may have important implications for developing therapeutic drugs.
Simpson
L L
LL
Department of Medicine, Jefferson Medical College, Philadelphia, Pennsylvania.
Coffield
J A
JA
Bakry
N
N
D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors*; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001905:Botulinum Toxins / Q000037:antagonists & inhibitors* / Q000494:pharmacology; D004347:Drug Interactions; D066298:In Vitro Techniques; D007425:Intracellular Membranes / Q000201:enzymology; D007700:Kinetics; D018942:Macrolides; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D051379:Mice; D009045:Motor Endplate / Q000187:drug effects / Q000502:physiology; D009469:Neuromuscular Junction / Q000187:drug effects / Q000502:physiology; D009498:Neurotoxins / Q000037:antagonists & inhibitors* / Q000494:pharmacology; D010741:Phospholipases A / Q000187:drug effects*; D054467:Phospholipases A2; D009435:Synaptic Transmission / Q000187:drug effects / Q000502:physiology; D013744:Tetanus Toxin / Q000037:antagonists & inhibitors; D014617:Vacuoles / Q000201:enzymology*
D000900:Anti-Bacterial Agents; D018942:Macrolides; D009498:Neurotoxins; D013744:Tetanus Toxin; C040929:bafilomycin A1; D010741:Phospholipases A; D054467:Phospholipases A2; D001905:Botulinum Toxins; D000251:Adenosine Triphosphatases