Tetanus Toxin Fragment C Binds to a Protein Present in Neuronal Cell Lines and Motoneurons

Herreros, Judit; Lalli, Giovanna; Montecucco, Cesare; Schiavo, Giampietro

doi:10.1046/j.1471-4159.2000.0741941.x

Cited by 79 publications

(29 citation statements)

References 52 publications

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“…Adrenal chromaffin cells, after the addition of exogenous ganglioside, are intoxicated by TeNT, which blocks evoked release of catecholamine (36). Differentiated PC12 cells, a rat pheochromocytoma cell line, internalize HCR/T and TeNT (7,37) mediated by the C-terminal subdomain of the HCR (38). The intoxication of PC12 cells by TeNT is dependent on the presence of cholesterol (10), but a further mechanism for TeNT entry has not been defined.…”

Section: Discussionmentioning

confidence: 99%

Entry of a Recombinant, Full-Length, Atoxic Tetanus Neurotoxin into Neuro-2a Cells

et al. 2014

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Tetanus neurotoxin (TeNT) and botulinum neurotoxin (BoNT) are clostridial neurotoxins (CNTs) that are the most toxic proteins for humans (1). TeNT and BoNT share ϳ35% identity and ϳ65% similarity and overall structure-function properties (2). BoNT intoxication results in flaccid paralysis through the inhibition of acetylcholine release by motor neurons, while TeNT intoxication yields a spastic paralysis due to inhibition of glycine release by inhibitory neurons (3). TeNT and BoNT are expressed as ϳ150-kDa single-chain proteins that are cleaved to form dichain proteins linked by a disulfide bond (2). The N-terminal 50-kDa light chain (LC) is a zinc-metalloprotease that cleaves neuronspecific soluble NSF attachment protein (SNAP) receptor (SNARE) proteins (4). TeNT and BoNT serotype B cleave the same residue within vesicle-associated membrane protein 2 (VAMP2), a SNARE protein of synaptic vesicles (SVs). The C-terminal 100-kDa heavy chain (HC) contains two structurally distinct domains with separate functions. The translocation domain (HCT) facilitates LC translocation from the SV lumen into the cell cytosol, and the receptor binding domain (HCR) binds dual host receptors.BoNT/A binds a ganglioside and synaptic vesicle protein 2 (SV2) and enters neurons upon SV recycling from the plasma membrane (5). Upon SV acidification within the periphery of motor neurons, the HCT undergoes a pH-dependent conformational change and inserts into the SV membrane, forming a channel that allows the LC to escape into the cytosol. Within the periphery of the motor neuron, the LC cleaves SNARE proteins, resulting in loss of stimulatory signaling between neurons and muscles, yielding flaccid paralysis. The LC of BoNT/A localizes to the plasma membrane to target synaptosomal-associated protein 25 (SNAP25) for cleavage within neurons (6).TeNT binds two gangliosides as functional receptors (7). TeNT can bind a glycophosphatidylinositol (GPI)-anchored protein (8) or SV2 (9), but the significance of these interactions has not been defined (10) or reproduced (11), respectively. TeNT enters motor neurons upon endocytosis (12) and traffics through motor neurons associated with Rab7-enriched endosomes that are of neutral pH (13,14). Retrograde trafficking proceeds from the axon to the soma, where TeNT transcytoses from the motor neuron into an inhibitory neuron of the central nervous system (CNS). Upon vesicle acidification, the LC is translocated into the cytosol and cleaves VAMP2. The block in signaling between the inhibitory neurons and motor neurons leads to the spastic paralysis characteristic of tetanus. The molecular mechanism responsible for the unique entry of BoNT and TeNT is not clearly understood.The modular structural domains of the CNTs have permitted the study of individual domains to assess protein structure-function in vitro; the LC is a functional SNARE protease (15), the HCT inserts into lipid bilayers and forms a channel (16), and the HCR binds host receptors (17). The tetanus HCR (HCR/T) can retrograde traffic in both cultu...

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Section: Discussionmentioning

confidence: 99%

Entry of a Recombinant, Full-Length, Atoxic Tetanus Neurotoxin into Neuro-2a Cells

et al. 2014

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“…The fragment TTC is necessary (and sufficient) for neuron binding and internalisation, retrograde and transynaptic transport [18,19]. Moreover, studies both in vitro and in vivo demonstrated that TTC has the capacity to deliver other large proteins linked by chemical or genetic fusion such as horseradish perodixase [20], b-galactosidase [17,21], superoxide dismutase [22][23][24], cardiotrophin-1 [25] and survival motor neuron factor in neurons [26].…”

Section: Introductionmentioning

confidence: 99%

Antiapoptotic activity maintenance of Brain Derived Neurotrophic Factor and the C fragment of the tetanus toxin genetic fusion protein

Ciriza¹,

García‐Ojeda

Martín-Burriel³

et al. 2008

Open Life Sciences

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Neurotrophic factors have been widely suggested as a treatment for multiple diseases including motorneuron pathologies, like Amyotrophic Lateral Sclerosis. However, clinical trials in which growth factors have been systematically administered to Amyotrophic Lateral Sclerosis patients have not been effective, owing in part to the short half-life of these factors and their low concentrations at target sites. A possible strategy is the use of the atoxic C fragment of the tetanus toxin as a neurotrophic factor carrier to the motorneurons.The activity of trophic factors should be tested because their genetic fusion to proteins could alter their folding and conformation, thus undermining their neuroprotective properties. For this purpose, in this paper we explored the Brain Derived Neurotrophic Factor (BDNF) activity maintenance after genetic fusion with the C fragment of the tetanus toxin. We demonstrated that BDNF fused with the C fragment of the tetanus toxin induces the neuronal survival Akt kinase pathway in mouse cortical culture neurons and maintains its antiapoptotic neuronal activity in Neuro2A cells.

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“…Despite variations within their targets and biological activities, CNT share a common structure composed of a 50 kDa light (LC) and a 100 kDa heavy chain (HC) linked by a single disulfide bond . The 100 kDa HC mediates the binding and internalization of TeNT into neurons, while the 50 kDa LC is a zinc metalloprotease containing a catalytic domain of the neurotoxin that inhibits neurotransmission (Herreros et al, 2000).…”

Section: Introductionmentioning

confidence: 99%

“…Structural analysis of HC revealed that the fragment C binding domain is composed of two sub-domains: the proximal H CN subdomain and the extreme carboxyl sub-domain, H CC . Several lines of evidence suggest that a set of conserved residues within the H CC sub-domain form the ganglioside binding motif and take part in neuron binding activity of TeNT (Herreros et al, 2000;Turton et al, 2002).…”

Section: Introductionmentioning

confidence: 99%

Comparative human and mouse antibody responses against tetanus toxin at clonal level

Yousefi

Younesi

Bayat

et al. 2015

Journal of Immunotoxicology

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Tetanus is a highly fatal disease caused by tetanus neurotoxin (TeNT) and remains a major threat to human and animal health, despite preventive strategies. TeNT is composed of heavy and light chain linked by a disulfide bond. The antibody response to TeNT is polyclonal and directed to multiple epitopes within both the light and heavy chains, leading to toxin neutralization. This study was undertaken to localize and compare neutralizing epitopes recognized by human and mouse TeNT-specific antibodies at a clonal level. In the present study, 22 murine hybridoma clones and 50 human lymphoblastoid cell lines secreting monoclonal antibodies (mAb) were generated against TeNT. The specificity of these mAb was determined using different recombinant fragments of tetanus toxin. Moreover, this study investigated the in vitro toxin neutralizing activity of these mAb by a ganglioside GT1b assay. The results showed that tetanus toxoid immunization in humans and BALB/c mice induced a vigorous humoral immune response against different fragments of TeNT, particularly the carboxyl-terminal fragment of the heavy chain (known as fragment C). The fragment C-specific human and mouse mAb could largely neutralize TeNT. However, while all fragment C-specific human mAb reacted with the carboxyl-terminal part of this fragment (H CC ), the majority of the mouse mAb failed to recognize this region. These results suggested that fragment C is the major target for the TeNT neutralizing antibodies, although different epitopes seem to be targeted by human and mouse antibodies.

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Tetanus Toxin Fragment C Binds to a Protein Present in Neuronal Cell Lines and Motoneurons

Cited by 79 publications

References 52 publications

Entry of a Recombinant, Full-Length, Atoxic Tetanus Neurotoxin into Neuro-2a Cells

Entry of a Recombinant, Full-Length, Atoxic Tetanus Neurotoxin into Neuro-2a Cells

Antiapoptotic activity maintenance of Brain Derived Neurotrophic Factor and the C fragment of the tetanus toxin genetic fusion protein

Comparative human and mouse antibody responses against tetanus toxin at clonal level

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