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Advances in Clostridial and Related Neurotoxins, 3rd Edition

A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Molecular Toxicology".

Deadline for manuscript submissions: 20 May 2025 | Viewed by 1529

Special Issue Editor

Special Issue Information

Dear Colleagues,

Botulinum neurotoxins (BoNTs), the causative agents of the potentially lethal vertebrate disease botulism, comprise a large and expanding family of protein toxins produced by various bacterial strains of the genus Clostridium. BoNTs are significant as disease-causing agents, potential bioterrorist agents, and as unique, long-lasting, and widely used bio-pharmaceutical agents. Currently, BoNTs are categorized into seven immunologically distinct serotypes, with several subtypes within each serotype. However, in recent years, discoveries of novel BoNTs, as well as potential BoNT homologues in other organisms, have challenged this categorization and expanded the family of BoNTs. While novel BoNTs are continually being identified by sequencing, most have not been purified and functionally characterized. The further identification and characterization of novel and known BoNTs will yield insights into the evolutionary forces driving the diversity of this protein toxin family and potentially reveal as-yet-unknown pharmacologic properties of BoNTs, with the potential to lead to novel or improved BoNT-based biopharmaceuticals. Furthermore, genetic methods now allow for the construction of recombinant and chimeric BoNTs, enabling the directed engineering of BoNTs with defined amino acid or functional domain substitutions. Combined with ongoing structural analyses, these studies will lead to a deeper understanding of the molecular mechanisms underlying the toxicity and pharmacologic potential of a large family of BoNTs. Both approaches exploring novel BoNTs and recombinant studies are exciting avenues of research, with the potential to open the door to unlocking the underlying molecular and evolutionary mechanisms of the high potency of BoNTs, eventually leading to improved safety approaches, countermeasure development, and novel pharmaceuticals and pharmaceutical applications.

Dr. Sabine Pellett
Guest Editor

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Keywords

  • botulinum neurotoxin
  • BoNT
  • toxicity
  • molecular mechanisms
  • recombinant
  • derivative
  • clostridium botulinum

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Published Papers (1 paper)

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Research

18 pages, 2471 KiB  
Article
Potency Evaluations of Recombinant Botulinum Neurotoxin A1 Mutants Designed to Reduce Toxicity
by Polrit Viravathana, William H. Tepp, Marite Bradshaw, Amanda Przedpelski, Joseph T. Barbieri and Sabine Pellett
Int. J. Mol. Sci. 2024, 25(16), 8955; https://doi.org/10.3390/ijms25168955 - 17 Aug 2024
Cited by 1 | Viewed by 1155
Abstract
Recombinant mutant holotoxin BoNTs (rBoNTs) are being evaluated as possible vaccines against botulism. Previously, several rBoNTs containing 2–3 amino acid mutations in the light chain (LC) showed significant decreases in toxicity (2.5-million-fold–12.5-million-fold) versus wild-type BoNT/A1, leading to their current exclusion from the Federal [...] Read more.
Recombinant mutant holotoxin BoNTs (rBoNTs) are being evaluated as possible vaccines against botulism. Previously, several rBoNTs containing 2–3 amino acid mutations in the light chain (LC) showed significant decreases in toxicity (2.5-million-fold–12.5-million-fold) versus wild-type BoNT/A1, leading to their current exclusion from the Federal Select Agent list. In this study, we added four additional mutations in the receptor-binding domain, translocation domain, and enzymatic cleft to further decrease toxicity, creating 7M rBoNT/A1. Due to poor expression in E. coli, 7M rBoNT/A1 was produced in an endogenous C. botulinum expression system. This protein had higher residual toxicity (LD50: 280 ng/mouse) than previously reported for the catalytically inactive rBoNT/A1 containing only three of the mutations (>10 µg/mouse). To investigate this discrepancy, several additional rBoNT/A1 constructs containing individual sets of amino acid substitutions from 7M rBoNT/A1 and related mutations were also endogenously produced. Similarly to endogenously produced 7M rBoNT/A1, all of the endogenously produced mutants had ~100–1000-fold greater toxicity than what was reported for their original heterologous host counterparts. A combination of mutations in multiple functional domains resulted in a greater but not multiplicative reduction in toxicity. This report demonstrates the impact of production systems on residual toxicity of genetically inactivated rBoNTs. Full article
(This article belongs to the Special Issue Advances in Clostridial and Related Neurotoxins, 3rd Edition)
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