Botulinal neurotoxins: revival of an old killer
Introduction
Seven biochemically and serologically distinct botulinum neurotoxins (designated BoNT/A to/G) have been characterized from anaerobic spore-forming bacteria of the genus Clostridium. The bacterial spores are widely present in the environment and are resistant to various physical and chemical agents. Therefore, they can contaminate food and, under suitably anaerobic conditions, germinate and yield the vegetative bacterial cells that produce one or more of the toxins. Ingestion of BoNT-poisoned food causes an intoxication known as botulism, which was first described at the beginning of the 17th century and associated to a protein neurotoxin at the end of that same century [1, 2, 3]. More recently, infant botulism was described as a syndrome caused by the colonization of the intestinal tract of infants by neurotoxigenic strains of C. botulinum, which subsequently results in infant intoxication [2, 3]. Another form of the disease is wound botulism, which results from the production of BoNTs by C. botulinum growing in anaerobic wounds [2, 3].
This brief overview highlights the structural organization and mode of action of BoNTs on neuroexocytosis. The review also focuses on the therapeutic uses and current limitations of BoNTs, as well as the ongoing developments in their use.
Section snippets
Botulism
The signs and symptoms of botulism are essentially the same for all forms of the disease, and are a consequence of sustained blockade of acetylcholine (ACh) release at somatic and autonomic nerve terminals. The facial and throat muscles are the first skeletal muscles that become weak and progressively paralyzed, causing diplopia, ptosis, dysphagia and facial paralysis. The paralysis progressively descends to affect muscles of the trunk, including the respiratory and visceral muscles (if death
Structure of botulinal neurotoxins
Before their release via bacterial autolysis, BoNTs are produced as progenitor toxins; that is, as variously sized complexes composed of inactive single polypeptide toxin chains (Mr ∼150 kDa) and other non-toxic accessory proteins. The accessory proteins protect BoNTs during their passage through the stomach, and they dissociate from the progenitor toxins when the complexes reach the small intestine — the main site at which the BoNTs are absorbed into the bloodstream and subsequently reach their
Botulinal neurotoxin-elicited inhibition of neuroexocytosis
The BoNTs are the most potent toxins known. The mouse LD50 values for the BoNTs range from 1 ng/kg to 5 ng/kg, and similar or lower values have been estimated for humans [16]. Their toxicity is usually expressed in units: 1 unit is 1 mouse LD50 for a 20 g Swiss strain mouse, and the LD50 of BoNT/A for an adult human (∼ 70 kg) has been estimated to be ∼ 3000 units. This potency derives from their high neurospecificity and from their catalytic activity, which results in the blockade of essential
Therapeutic uses of botulinal neurotoxins
BoNT/A, under the trade names of BOTOX (Allergan) and Dysport (Ipsen), has become a multi-million dollar drug used in the treatment of many human syndromes and as a ‘pharmaco-cosmetic’ [18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33]; BoNT/B is also commercially available (Neurobloc, Elan Pharmaceuticals). The increasing use and commercial success of BoNTs is due to their high specificity for peripheral cholinergic nerve terminals and to the remarkable fact that, when proper
Current limitations of the therapeutic use of botulinal neurotoxins
As mentioned above, the effect of BoNTs is reversible; therefore, to sustain the desired therapeutic effect, treatments usually have to be repeated. Thus, as BoNTs are antigenic, an immune response leading to the formation of neutralizing antibodies can occur [47, 48]. This possibility is rare because most treatments require only minute amounts of BoNT/A; however, it could become a problem when larger amounts of toxin are needed to treat powerful muscles of body regions rich in lymph nodes
Conclusions
In conclusion, BoNTs are zinc metalloendoproteases that exhibit extraordinary specificities for proteins involved in the neurotransmitter release process, and their toxicities make them responsible for animal and human botulism. However, these extremely poisonous molecules have become useful therapeutic agents in a number of expanding applications in human medicine. Ongoing efforts to elucidate BoNTs’ mechanisms of action for reducing pain and other pathological conditions will provide an
References and recommended reading
Papers of particular interest, published within the annual period of review, have been highlighted as:
• of special interest
•• of outstanding interest
Acknowledgements
We apologize to our colleagues whose papers could not be cited owing to a restriction on space. We thank Dr Arnold S. Kreger for helpful suggestions during the copyediting of the manuscript. The authors’ studies of BoNTs were supported by research grants from Telethon-Italia Grant GP0272/01 (to CM) and from the Direction des Systèmes de Forces et de la Prospective (Grant # 026065093 to JM).
References (56)
- et al.
Presynaptic receptor arrays for clostridial neurotoxins
Trends Microbiol
(2004) - et al.
Sequence homology and structural analysis of the clostridial neurotoxins
J Mol Biol
(1999) - et al.
The journey of tetanus and botulinum neurotoxins in neurons
Trends Microbiol
(2003) - et al.
The high-affinity binding of Clostridium botulinum type B neurotoxin to synaptotagmin II associated with gangliosides GT1b/GD1a
FEBS Lett
(1996) - et al.
Synaptotagmin I and II act as nerve cell receptors for botulinum neurotoxin G
J Biol Chem
(2004) - et al.
Membrane fusion
Cell
(2003) - et al.
Dynamics of motor nerve terminal remodelling unveiled using SNARE-cleaving botulinum toxins: the extent and duration are dictated by the sites of SNAP-25 truncation
Mol Cell Neurosci
(2003) - et al.
Pharmacokinetic properties of different formulations of botulinum neurotoxin type A
Mov Disord
(2004) Use of botulinum toxin in neuro-ophthalmology
Curr Opin Ophthalmol
(2001)- Hyperhydrosis and botulinum toxin in dermatology. Edited by Kreyden OP, Boni R, Burg G. Basel: Karger;...
Management of Frey's syndrome and hypersialorrhea with botulinum toxin
Facial Plast Surg Clin North Am
Botulinum toxin and other new approaches to migraine therapy
Annu Rev Med
Subcutaneous administration of botulinum toxin A reduces formalin-induced pain
Pain
Regulation of calcitonin gene-related peptide secretion from trigeminal nerve cells by botulinum toxin type A: implications for migraine therapy
Headache
Antiepileptic effects of botulinum neurotoxin E
J Neurosci
Botulinum neurotoxin serotypes A and C do not affect motor units survival in humans: an electrophysiological study by motor units counting
Clin Neurophysiol
Clinical usefulness of botulinum toxin in the lower extremity
Foot Ankle Clin
Historical aspects of botulinum toxin: Justinus Kerner (1786-1862) and the “sausage poison”
Neurology
Neurotoxins affecting neuroexocytosis
Physiol Rev
Indentification of the major steps in botulinum toxin action
Annu Rev Pharmacol Toxicol
Structural analysis of the catalytic and binding sites of Clostridium botulinum neurotoxin B
Nat Struct Biol
The Hcc-domain of botulinum neurotoxin A and B exhibit a singular ganglioside binding site displaying serotype specific carbohydrate interaction
Mol Microbiol
Synaptotagmins I and II mediate entry of botulinum neurotoxin B into cells
J Cell Biol
Truncated SNAP-25 (1-197), like botulinum neurotoxin A, can inhibit insulin secretion from HIT-T15 insulinoma cells
Mol Endocrinol
A single amino acid near the C terminus of the synaptosome-associated protein of 25 kDa (SNAP-25) is essential for exocytosis in chromaffin cells
Proc Natl Acad Sci USA
Bacterial toxins. A table of lethal amounts
Microbiol Rev
Evaluation of the therapeutic usefulness of botulinum neurotoxin B, C, E and F compared with the long lasting type A. Basis for distinct duration of inhibition of exocytosis in central neurons
J Biol Chem
Development of botulinum toxin therapy
Dermatol Clin
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