Structural flexibility of the tetanus neurotoxin revealed by crystallographic and solution scattering analyses

Zhang, Chun-ming; Imoto, Yoshihiro; Hikima, Takaaki; Inoue, Tsuyoshi

doi:10.1016/j.yjsbx.2021.100045

Cited by 6 publications

(5 citation statements)

References 50 publications

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“…Recent studies on anti-TeNT humAbs reported that the highest TeNT neutralization was displayed by antibodies recognizing HC, particularly those preventing PSG binding, but did not report on HC-N recognition (39)(40)(41)(42). Although the role of HC-N in neuron intoxication is currently unknown, its deletion causes loss of toxicity (15).…”

Section: Humabs Recognize Distinct Domains Of Tent and Display Distinct Neutralizing Capabilitymentioning

confidence: 99%

“…The isolation of anti-TeNT humAbs from human B memory cells was recently reported (39)(40)(41)(42). TeNT neutralization in mice could be achieved only by preincubation of the toxin with a combination of 3 antibodies and/or using high molar excess of humAbs with respect to TeNT.…”

Section: The Structure Of the [Tent]-[tt104-fab]-[tt110-fab] Trimeric Immunocomplex Has Been Resolvedmentioning

confidence: 99%

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Exceptionally potent human monoclonal antibodies are effective for prophylaxis and treatment of tetanus in mice

Pirazzini¹,

Grinzato²,

Corti³

et al. 2021

Journal of Clinical Investigation

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Human monoclonal antibodies were used here to study the mechanism of neuron intoxication by tetanus neurotoxin and to evaluate them as a safe preventive and therapeutic substitute of hyperimmune sera for tetanus in mice. By screening memory B cells of immune donors, we selected two monoclonal antibodies specific for tetanus neurotoxin with exceptionally high neutralizing activities, which were extensively characterized both structurally and functionally. We found that these antibodies interfere with the binding and translocation of the neurotoxin into neurons by interacting with two epitopes, whose definition pinpoints crucial events in the cellular pathogenesis of tetanus. This information explains the unprecedented neutralization ability of these antibodies, which were found to be exceptionally potent in preventing experimental tetanus when injected in mice long before the neurotoxin. Moreover, their Fab derivatives neutralized tetanus neurotoxin in post-exposure experiments, suggesting their potential therapeutic use via intrathecal injection. As such, these human monoclonal antibodies, as well as their Fab derivatives, meet all requirements for being considered for prophylaxis and therapy of human tetanus and are ready for clinical trials.

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Section: Humabs Recognize Distinct Domains Of Tent and Display Distinct Neutralizing Capabilitymentioning

confidence: 99%

Section: The Structure Of the [Tent]-[tt104-fab]-[tt110-fab] Trimeric Immunocomplex Has Been Resolvedmentioning

confidence: 99%

Exceptionally potent human monoclonal antibodies are effective for prophylaxis and treatment of tetanus in mice

Pirazzini¹,

Grinzato²,

Corti³

et al. 2021

Journal of Clinical Investigation

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“…Some of the X-ray crystallographic structures have been determined at acidic pH values in the range 4-6 [8], but no structural variation has been observed. Interestingly, BoNTs are closely related to the tetanus toxin TeNT, which presents a closed conformation with different organization, when compared to BoNT/E1 [11,12]. BoNT/A associates with the non-toxic non-hemagglutinin (NTNH) protein at acidic pH, forming stable complexes resistant to protease and acidic degradation [13].…”

Section: Introductionmentioning

confidence: 99%

In Silico Conformational Features of Botulinum Toxins A1 and E1 According to Intraluminal Acidification

Cottone

Chiodo

Maragliano

et al. 2022

Toxins

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Although botulinum neurotoxins (BoNTs) are among the most toxic compounds found in nature, their molecular mechanism of action is far from being elucidated. A key event is the conformational transition due to acidification of the interior of synaptic vesicles, leading to translocation of the BoNT catalytic domain into the neuronal cytosol. To investigate these conformational variations, homology modeling and atomistic simulations are combined to explore the internal dynamics of the sub-types BoNT/A1 (the most-used sub-type in medical applications) and BoNT/E1 (the most kinetically efficient sub-type). This first simulation study of di-chain BoNTs in closed and open states considers the effects of both neutral and acidic pH. The conformational mobility is driven by domain displacements of the ganglioside-binding site in the receptor binding domain, the translocation domain (HCNT) switch, and the belt α-helix, which present multiple conformations, depending on the primary sequence and the pH. Fluctuations of the belt α-helix are observed for closed conformations of the toxins and at acidic pH, while patches of more solvent-accessible residues appear under the same conditions in the core translocation domain HCNT. These findings suggest that, during translocation, the higher mobility of the belt could be transmitted to HCNT, leading to the favorable interaction of HCNT residues with the non-polar membrane environment.

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“…The spatial localizations of the epitopes within the structure of TeNT were mapped to the crystallographic model [45,46] available in PDB (PDB: 1xdt) (Figure 4A), which displays the spatial location of the forty-three reactive epitopes identified by the SPOT synthesis array experiments (Table 1). Thirteen of the identified epitopes were exclusively in loop/coil structures while eleven shared amino acids in coil and helix or ß-sheet (Table 1 and Figure 4).…”

Section: The Spatial Location Of Tent Reactive Epitopesmentioning

confidence: 99%

High-Throughput IgG Epitope Mapping of Tetanus Neurotoxin: Implications for Immunotherapy and Vaccine Design

De-Simone¹,

Napoleão-Pêgo²,

Lechuga³

et al. 2023

Preprint

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Background: Tetanus is an acute, fatal disease caused by exotoxins released from Clostridium tetani during infections. A protective humoral immune response can be induced by vaccinations with pediatric and booster combinatorial vaccines that contain inactivated tetanus neurotoxin (TeNT) as a major antigen. Although some epitopes in NeNT have been described using various ap-proaches, a comprehensive list of its antigenic determinants that are involved with immunity have not been elucidated. To that end, a high resolution analysis of the linear B-cell epitopes in NeNT was performed using antibodies generated in vaccinated children. Methods: Two hundred sixty-four peptides that cover the entire coding sequence of TeNT protein were prepared in situ on a cellulose membrane by SPOT synthesis and probed with sera from children vaccinated (ChVS) with a triple DTP-vaccine to map continuous B-cell epitopes that were further characterized and validated by immunoassays. Results: Forty-three IgG epitopes were identified. Four (TT-215-218) were chemically synthesized as Multiple Antigen Peptides (MAPs) and used in peptide ELISAs to screen post-pandemic DTP vaccinations. The assay displayed a high performance with high sensitivity (99.99%) and specificity (100%). Conclusions: The complete map of linear IgG epitopes induced by vaccination with inactivated TeNT highlights three key epitopes involved in the ef-ficacy of the vaccine. Antibodies against epitope TT-8/G can block enzymatic activity, and those against epitopes TT-40/G and TT-42/G can interfere with TeNT binding to neuronal cell receptors. We further show that four of the epitopes identified can be employed in peptide ELISAs to assess vaccine coverage. Overall, the data suggest a set of select epitopes to engineer new, directed vaccines

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Structural flexibility of the tetanus neurotoxin revealed by crystallographic and solution scattering analyses

Abstract: Graphical abstract

Cited by 6 publications

References 50 publications

Exceptionally potent human monoclonal antibodies are effective for prophylaxis and treatment of tetanus in mice

Exceptionally potent human monoclonal antibodies are effective for prophylaxis and treatment of tetanus in mice

In Silico Conformational Features of Botulinum Toxins A1 and E1 According to Intraluminal Acidification

High-Throughput IgG Epitope Mapping of Tetanus Neurotoxin: Implications for Immunotherapy and Vaccine Design

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