Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.4.24.69 (botulinum neurotoxin)
 1,901 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Two DNA fragments, 3 kbp and 7.8kbp, which encode the type C1 botulinum neurotoxin gene, were obtained from toxigenic bacteriophage DNA by treatment with a restriction enzyme. They were cloned into the plasmid vectors for nucleotide sequence determination. The nucleotide sequence contained a single open reading frame coding for 1,291 amino acids corresponding to a polypeptide with a molecular weight of 149,000. The amino acid sequence of the C1 toxin has a few regions highly homologous with tetanus toxin.
 ...
PMID:The complete nucleotide sequence of the gene coding for botulinum type C1 toxin in the C-ST phage genome. 222 45

Injection of exogenous mRNA purified from various tissue preparations into cellular translation systems such as Xenopus oocytes has allowed expression of complex proteins (e.g., receptors for neurotransmitters). No evidence for expression of injected exogenous mRNA, however, has been reported in terminally differentiated neurons. If achieved, it would allow the study of long-lasting changes of properties of nerve cells in their functional context. To obtain evidence of such expression, we chose two proteins that produce a detectable effect even at very low intracellular concentrations. Tetanus toxin and botulinum neurotoxin fulfill this criterion, being the most potent neurotoxins known. Both toxins block neurotransmitter release at nanomolar intracellular concentrations. These di-chain proteins, consisting of a light chain and a heavy chain, have recently been sequenced. Their active sites are located (or partly located) on the light chain. mRNAs encoding the light chain of either toxin were transcribed in vitro from the cloned and specifically truncated genes of Clostridium tetani and Clostridium botulinum, respectively, and injected into presynaptic cholinergic neurons of the buccal ganglia of Aplysia californica. Depression of neurotransmitter release appeared in less than 1 hr, demonstrating successful expression of foreign mRNA injected into a neuron in situ.
 ...
PMID:Exogenous mRNA encoding tetanus or botulinum neurotoxins expressed in Aplysia neurons. 223

Experiments have been conducted that deal with the structure and biological activity of clostridial toxins. Studies have dealt mainly with botulinum neurotoxin, but work has also been done with tetanus toxin and with the binary toxin. Structural studies indicate that proteolytic processing of botulinum neurotoxin induces two major outcomes: activation and aging. The first is associated with a marked increase in toxicity and with conversion from a single chain to a dichain structure. The second is associated with nominal changes in toxicity and with molecular rearrangements in the dichain structure. Immunological studies have resulted in isolation and characterization of a monoclonal antibody that neutralizes tetanus toxin. Monoclonal antibodies have also been raised against botulinum neurotoxin, and these antibodies have been used to demonstrate that: i) activation is not due to marked conformational changes in the relevant epitopes, ii) binding of the toxin to cholinergic nerve endings does not produce detectable conformational changes, and iii) all functional domains of the toxin appear to be internalized simultaneously. Immunological studies done in vivo and in vitro suggest that certain antibodies may enter cholinergic nerves and neutralize subsequently internalized toxin. Additional work on clostridial toxins has produced the following results: i) the ligand binding assay typically used with tetanus toxin (i.e., low pH and ionic strength) is of questionable biological significance, ii) the binary toxin, like the clostridial neurotoxins, enters cells by receptor-mediated endocytosis, and iii) tetanus toxin can alter the disposition of protein kinase C in one neuroblastoma cell line.
 ...
PMID:The study of clostridial and related toxins. The search for unique mechanisms and common denominators. 229 Jan 29

Tetanus and botulinum neurotoxins (TeNT and BoNT) bind strongly and specifically to the nervous tissue, as it can be inferred from their potency and from their effects restricted to the nervous system. The molecular basis of these properties are presently unknown. As a first approach, we have investigated the interaction of TeNT and BoNT with model membranes by photolabelling with phospholipid analogues carrying the photoreceptor group at different positions of the lipid molecule in order to probe different membrane regions. We found that at neutral pH TeNT and BoNTs (type A, B and E) adsorb onto the surface of negatively charged liposomes. Polysialogangliosides increase this interaction only slightly thus suggesting that they provide a minor contribution to toxin lipid binding. On this basis we propose that clostridial neurotoxins bind to lipids via both a predominant unspecific interaction with negatively charged lipids (including gangliosides) and a specific, but weaker, interaction with polysialogangliosides. At acidic pH values both chains of these neurotoxins are labelled strongly by photogroups located in the hydrophobic milieu of the membrane with a pH dependence that overlaps the range of pH values reached in the endosomal lumen. This result is consistent with their insertion into the lipid bilayer in agreement with the idea that clostridial neurotoxins may penetrate into cells via intracellular low pH compartments.
 ...
PMID:Membrane interactions of tetanus and botulinum neurotoxins: a photolabelling study with photoactivatable phospholipids. 229 Jan 32

Permeabilisation of PC12 cells with digitonin allowed a direct study of the intracellular action of botulinum neurotoxin A, one of a group of dichain proteins produced by Clostridium botulinum that causes the fatal neuroparalytic condition, botulism. Release of [3H]noradrenaline from these permeabilised cells could be evoked by Ca2+ and this was inhibited specifically by the neurotoxin in a dose-dependent manner (half-maximal dose approximately 2 nM under the conditions used). Inclusion of the reducing agent dithiothreitol (up to 10 mM) had no effect on the level of inhibition. Moreover, electrophoretic analysis showed that this treatment of the toxin in the native state caused negligible reduction of inter-chain disulphide bonds. Toxin-induced blockade of neurotransmitter release was incomplete and could not be overcome by increased Ca2+ concentration (100 microM). The observed toxin-insensitivity of the release from intact PC12 cells must result from inefficient toxin uptake, relative to that in peripheral cholinergic neurones. Refolded light chain alone inhibited exocytosis to the same degree and with similar potency to that of the intact neurotoxin, an effect not altered by the heavy chain. This inhibitory activity of the light chain in PC12 cells accords with observations made in permeabilised chromaffin cells [(1989) J. Biol. Chem. 264, 10354-10360; (1989) FEBS Lett. 255, 391-394] but contrasts with invertebrate neurones, where intracellular injection of the same preparations of both chains were necessary for inhibition of quantal release of acetylcholine [(1988) Proc. Natl. Acad. Sci. USA 85, 4090-4094]. These collective findings may signify an interesting difference in the release process in such diverse systems or denote a dissimilarity in the transport or processing of the toxin when applied into intact neurones or cells permeabilised by detergent or streptolysin.
 ...
PMID:Ca2(+)-dependent noradrenaline release from permeabilised PC12 cells is blocked by botulinum neurotoxin A or its light chain. 231 61

1. Tacrine (20 microM) induced, like 4-aminoquinoline (4-AQ, 200 microM), the appearance of a population of miniature endplate potentials (m.e.p.ps) with more than twice the normal amplitude or time-to-peak. The times-to-peak of nerve impulse-evoked endplate potentials were not similarly affected. 2. Cholinesterase inhibition by edrophonium (25 microM) did not prevent tacrine or 4-AQ from inducing this population of m.e.p.ps. 3. Nerve-muscle preparations in which the normal calcium-sensitive quantal release of acetylcholine had been blocked by botulinum neurotoxin type A also responded to tacrine by an increase in the frequency of giant or slow m.e.p.ps. 4. Reduction of the temperature from 30 degrees to 14 degrees C reduced the frequency of giant or slow m.e.p.ps induced either by tacrine or by 4-AQ. A similar effect was obtained by colchicine (5 mM). This supports the idea that proximo-distal axonal transport is required for the secretory activity. 5. The neurosecretion evoked by tacrine could explain the therapeutic effects of the drug claimed in the treatment of Alzheimer's type of dementia.
 ...
PMID:Tetrahydroaminoacridine (tacrine) stimulates neurosecretion at mammalian motor endplates. 239 Jun 74

The interaction of botulinum neurotoxin (Botx) with planar lipid membranes was studied by measuring the ability of the toxin to form ion-conducting channels. Channel formation was pH dependent. At physiological pH, Botx formed no channels, whereas at pH 6.6, the toxin formed channels with a unit conductance of 12 pS in 0.1 M NaCl. The rate of channel formation increased with decreasing pH, reaching a maximum at pH 6.1, and then decreased at lower values of pH. The channels, once formed, were permanent entities in the membrane throughout the course of an experiment and fluctuated between an open and a closed state. The rate of channel formation depended upon the square of the toxin concentration, suggesting an aggregation step is involved in channel formation. The data were consistent with the hypothesis that Botx enters cells through endocytosis, followed by its release into the cytoplasm at low pH.
 ...
PMID:Ion-conducting channels produced by botulinum toxin in planar lipid membranes. 242 93

The exocytotic release of L-glutamate from guinea-pig cerebral cortical synaptosomes can be extensively inhibited by preincubation with botulinum neurotoxin type A at 37 degrees C for 1-2 h. The toxin has no effect on synaptosomal respiratory control, respiratory capacity, ATP synthesis, plasma-membrane 86Rb+ permeability or plasma-membrane potential, does not inhibit the entry of 45Ca2+ into the synaptosome upon depolarization and does not alter the ability of intrasynaptosomal mitochondria to sequester Ca2+. The blockade of Ca2+-dependent glutamate release may be totally reversed by the Ca2+/2 H+-exchange ionophore ionomycin, but not by increasing extracellular Ca2+ concentration. It is suggested (a) that exocytosis is triggered by the penetration of Ca2+ into an intracellular hydrophobic milieu; (b) that this stage is blocked by the toxin and (c) that ionomycin is able to bypass this block and deliver Ca2+ to the exocytotic apparatus.
 ...
PMID:Botulinum toxin A blocks glutamate exocytosis from guinea-pig cerebral cortical synaptosomes. 243 34

1. A 50-kDa fragment representing the NH2-terminus of the heavy subunit of botulinum type A neurotoxin was found, at low pH, to evoke the release of K+ from lipid vesicles loaded with potassium phosphate. Similar K+ release was also observed with the intact neurotoxin, its heavy chain and a fragment consisting of the light subunit linked the 50-kDa NH2-terminal heavy chain fragment. The light subunit alone, however, was inactive. 2. In addition to K+, the channels formed in lipid bilayers by botulinum neurotoxin type A or the NH2-terminal heavy chain fragment were found to be large enough to permit the release of NAD (Mr 665). 3. The optimum pH for the release of K+ was found to be 4.5. Above this value K+ release rapidly decreased and was undetectable above pH 6.0. 4. The binding of radiolabelled botulinum toxin to a variety of phospholipids was assessed. High levels of toxin binding were only observed to lipid vesicles with an overall negative charge; much weaker binding occurred to lipid vesicles composed of electrically neutral phospholipids. 5. A positive correlation between the efficiency of toxin-binding and the efficiency of K+ release from lipid vesicles was not observed. Whereas lipid vesicles containing the lipids cardiolipin or dicetyl phosphate bound the highest levels of neurotoxin, the toxin-evoked release of K+ was low compared to vesicles containing either phosphatidyl glycerol, phosphatidyl serine or phosphatidyl inositol. 6. The implications of these observations to the mechanism by which the toxin molecule is translocated into the nerve ending are discussed.
 ...
PMID:A 50-kDa fragment from the NH2-terminus of the heavy subunit of Clostridium botulinum type A neurotoxin forms channels in lipid vesicles. 244 87

The pharmacologic activity of several clostridial neurotoxins was assayed on the mouse phrenic nerve-hemidiaphragm preparation. The substances that were assayed included botulinum neurotoxin types A, B, C and E and tetanus toxin. Experiments were done in the presence or absence of antagonists that inhibit either the internalization of toxins or intracellular expression of toxicity. Ammonium chloride and methylamine hydrochloride, agents that inhibit toxins that enter cells by receptor-mediated endocytosis, antagonized botulinum and tetanus neurotoxins. The magnitude of antagonism was substantial for all toxins. Calcium, 3,4-diaminopyridine and guanidine, agents that alter the intracellular expression of toxicity, produced a variable result. They were effective antagonists of botulinum neurotoxin type A, but they were less effective or inactive against the other neurotoxins. The ability of 3,4-diaminopyridine and guanidine to antagonize botulinum neurotoxin type A was highly calcium dependent. When ambient levels of the cation were reduced from 1.8 to 1.0 mM, the activity of the drugs was substantially reduced. The ability of these drugs to produce antagonism was also time dependent. When added simultaneously with toxin, they were maximally active; when added at later times, activity was diminished. A host of agents that alter intracellular levels of cyclic AMP, including theophylline, forskolin, isobutylmethylxanthine and cholera toxin, were evaluated as potential neurotoxin antagonists. Theophylline and isobutylmethylxanthine produced a transient increase in nerve-evoked muscle twitch. None of the drugs that alter tissue levels of cyclic AMP had a universal effect in antagonizing clostridial toxins. The data here have been compared with published data on drugs that antagonize binding of botulinum toxin and tetanus toxin.(ABSTRACT TRUNCATED AT 250 WORDS)
 ...
PMID:Use of pharmacologic antagonists to deduce commonalities of biologic activity among clostridial neurotoxins. 245 38


<< Previous 1 2 3 4 5 6 7 8 9 10 Next >>