Tetanus toxin is a zinc protein and its inhibition of neurotransmitter release and protease activity depend on zinc.
Tetanus and botulinum neurotoxins are the most potent toxins known. They bind to nerve cells, penetrate the cytosol and block neurotransmitter release. Comparison of their predicted amino acid sequences reveals a highly conserved segment that contains the HexxH zinc binding motif of metalloendopeptidases. The metal content of tetanus toxin was then measured and it was found that one atom of zinc is bound to the light chain of tetanus toxin. Zinc could be reversibly removed by incubation with heavy metal chelators. Zn2+ is coordinated by two histidines with no involvement in cysteines, suggesting that it plays a catalytic rather than a structural role. Bound Zn2+ was found to be essential for the tetanus toxin inhibition of neurotransmitter release in Aplysia neurons injected with the light chain. The intracellular activity of the toxin was blocked by phosphoramidon, a very specific inhibitor of zinc endopeptidases. Purified preparations of light chain showed a highly specific proteolytic activity against synaptobrevin, an integral membrane protein of small synaptic vesicles. The present findings indicate that tetanus toxin, and possibly also the botulinum neurotoxins, are metalloproteases and that they block neurotransmitter release via this protease activity.
Schiavo
G
G
Centro CNR Biomembrane, Università di Padova, Italy.
Poulain
B
B
Rossetto
O
O
Benfenati
F
F
Tauc
L
L
Montecucco
C
C
D000109:Acetylcholine / Q000378:metabolism*; D000200:Action Potentials / Q000187:drug effects; D000595:Amino Acid Sequence; D000818:Animals; D001048:Aplysia; D002540:Cerebral Cortex / Q000201:enzymology*; D005724:Ganglia / Q000187:drug effects / Q000502:physiology*; D066298:In Vitro Techniques; D008565:Membrane Proteins / Q000037:antagonists & inhibitors*; D008666:Metalloendopeptidases / Q000737:chemistry / Q000494:pharmacology*; D051379:Mice; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000037:antagonists & inhibitors*; D009474:Neurons / Q000187:drug effects / Q000502:physiology*; D010455:Peptides / Q000138:chemical synthesis / Q000494:pharmacology; D011480:Protease Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D011487:Protein Conformation; D050683:R-SNARE Proteins; D051381:Rats; D013054:Spectrophotometry, Atomic; D013329:Structure-Activity Relationship; D013569:Synapses / Q000187:drug effects / Q000502:physiology*; D013744:Tetanus Toxin / Q000737:chemistry / Q000494:pharmacology* / Q000633:toxicity; D015032:Zinc / Q000032:analysis / Q000378:metabolism / Q000494:pharmacology*
D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D010455:Peptides; D011480:Protease Inhibitors; D050683:R-SNARE Proteins; D013744:Tetanus Toxin; D008666:Metalloendopeptidases; C093556:zinc-endopeptidase, tetanus neurotoxin; D015032:Zinc; D000109:Acetylcholine