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TitleCryo-EM Structure Guided Engineering of Botulinum Neurotoxin A With Advanced Receptor Binding Affinity and Therapeutical Benefits.
Journal, issue, pagesAdv Sci (Weinh), Page e16713, Year 2026
Publish dateApr 7, 2026
AuthorsWenrui Wang / Zhaxi Zerang / Linjin You / Ziye Liu / Rong Nie / Fuwei Qi / Fenfen Gao / Chengmu Zhao / Wantong Ma / Jinghan He / Xiaoru Wang / Shanquan Wu / Bo Liu / Xinyao Liu / Dongsheng Lei / Dejuan Zhi / Dongsheng Wang /
PubMed AbstractBotulinum neurotoxin type A (BoNT/A) has been extensively used in treating a wide range of neurological disorders and aesthetics. However, insufficient binding affinity between BoNT/A and its ...Botulinum neurotoxin type A (BoNT/A) has been extensively used in treating a wide range of neurological disorders and aesthetics. However, insufficient binding affinity between BoNT/A and its receptor SV2C could lead to mild to severe side effects. An open active conformation of BoNT/A cryo-EM structure at a resolution of ∼2.85 Å was resolved. Guided by the BoNT/A cryo-EM structure, saturation mutagenesis libraries were constructed and screened by BACTH system to identify mutants that can boost toxin-SV2C affinity. The engineered toxin binding domain achieved up to approximately six-fold improved affinity in SPR analysis, and the engineered toxins exhibited significantly improved binding capacity, and enhanced SNAP25 cleavage efficacy in cultured neurons. Preclinical animal studies, including MLB, DAS, sweat test, and PET/CT assays, demonstrated that the engineered BoNT/A VLTS has higher potency, lower diffusion, and significantly better safety profiles than the BoNT/A wt, which can reduce side effects and benefit future therapeutic applications.
External linksAdv Sci (Weinh) / PubMed:41944347
MethodsEM (single particle)
Resolution2.8 - 2.85 Å
Structure data

51433

9gkq

EMDB-51433, PDB-9gkq:
Cryo-EM structure of Botulinum neurotoxin serotype A
Method: EM (single particle) / Resolution: 2.85 Å

64966

9vcs

EMDB-64966, PDB-9vcs:
Cryo-EM structure of semi-closed state of Botulinum neurotoxin serotype A at pH 7.4
Method: EM (single particle) / Resolution: 2.8 Å

Source
  • clostridium botulinum (bacteria)
  • clostridium botulinum (strain hall / atcc 3502 / nctc 13319 / type a) (bacteria)
KeywordsTOXIN / Botulinum neurotoxin serotype A / Cryo-EM

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