Cargando…

Structural characterisation of the catalytic domain of botulinum neurotoxin X - high activity and unique substrate specificity

Botulinum neurotoxins (BoNTs) are among the most potent toxins known and are also used to treat an increasing number of medical disorders. There are seven well-established serotypes (BoNT/A-G), which all act as zinc-dependent endopeptidases targeting specific members of the SNARE proteins required f...

Descripción completa

Detalles Bibliográficos
Autores principales: Masuyer, Geoffrey, Zhang, Sicai, Barkho, Sulyman, Shen, Yi, Henriksson, Linda, Košenina, Sara, Dong, Min, Stenmark, Pål
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5851995/
https://www.ncbi.nlm.nih.gov/pubmed/29540745
http://dx.doi.org/10.1038/s41598-018-22842-4
_version_ 1783306476003000320
author Masuyer, Geoffrey
Zhang, Sicai
Barkho, Sulyman
Shen, Yi
Henriksson, Linda
Košenina, Sara
Dong, Min
Stenmark, Pål
author_facet Masuyer, Geoffrey
Zhang, Sicai
Barkho, Sulyman
Shen, Yi
Henriksson, Linda
Košenina, Sara
Dong, Min
Stenmark, Pål
author_sort Masuyer, Geoffrey
collection PubMed
description Botulinum neurotoxins (BoNTs) are among the most potent toxins known and are also used to treat an increasing number of medical disorders. There are seven well-established serotypes (BoNT/A-G), which all act as zinc-dependent endopeptidases targeting specific members of the SNARE proteins required for synaptic vesicle exocytosis in neurons. A new toxin serotype, BoNT/X, was recently identified. It cleaves not only the canonical targets, vesicle associated membrane proteins (VAMP) 1/2/3 at a unique site, but also has the unique ability to cleave VAMP4/5 and Ykt6. Here we report the 1.35 Å X-ray crystal structure of the light chain of BoNT/X (LC/X). LC/X shares the core fold common to all other BoNTs, demonstrating that LC/X is a bona fide member of BoNT-LCs. We found that access to the catalytic pocket of LC/X is more restricted, and the regions lining the catalytic pocket are not conserved compared to other BoNTs. Kinetic studies revealed that LC/X cleaves VAMP1 with a ten times higher efficiency than BoNT/B and the tetanus neurotoxin. The structural information provides a molecular basis to understand the convergence/divergence between BoNT/X and other BoNTs, to develop effective LC inhibitors, and to engineer new scientific tools and therapeutic toxins targeting distinct SNARE proteins in cells.
format Online
Article
Text
id pubmed-5851995
institution National Center for Biotechnology Information
language English
publishDate 2018
publisher Nature Publishing Group UK
record_format MEDLINE/PubMed
spelling pubmed-58519952018-03-21 Structural characterisation of the catalytic domain of botulinum neurotoxin X - high activity and unique substrate specificity Masuyer, Geoffrey Zhang, Sicai Barkho, Sulyman Shen, Yi Henriksson, Linda Košenina, Sara Dong, Min Stenmark, Pål Sci Rep Article Botulinum neurotoxins (BoNTs) are among the most potent toxins known and are also used to treat an increasing number of medical disorders. There are seven well-established serotypes (BoNT/A-G), which all act as zinc-dependent endopeptidases targeting specific members of the SNARE proteins required for synaptic vesicle exocytosis in neurons. A new toxin serotype, BoNT/X, was recently identified. It cleaves not only the canonical targets, vesicle associated membrane proteins (VAMP) 1/2/3 at a unique site, but also has the unique ability to cleave VAMP4/5 and Ykt6. Here we report the 1.35 Å X-ray crystal structure of the light chain of BoNT/X (LC/X). LC/X shares the core fold common to all other BoNTs, demonstrating that LC/X is a bona fide member of BoNT-LCs. We found that access to the catalytic pocket of LC/X is more restricted, and the regions lining the catalytic pocket are not conserved compared to other BoNTs. Kinetic studies revealed that LC/X cleaves VAMP1 with a ten times higher efficiency than BoNT/B and the tetanus neurotoxin. The structural information provides a molecular basis to understand the convergence/divergence between BoNT/X and other BoNTs, to develop effective LC inhibitors, and to engineer new scientific tools and therapeutic toxins targeting distinct SNARE proteins in cells. Nature Publishing Group UK 2018-03-14 /pmc/articles/PMC5851995/ /pubmed/29540745 http://dx.doi.org/10.1038/s41598-018-22842-4 Text en © The Author(s) 2018 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Masuyer, Geoffrey
Zhang, Sicai
Barkho, Sulyman
Shen, Yi
Henriksson, Linda
Košenina, Sara
Dong, Min
Stenmark, Pål
Structural characterisation of the catalytic domain of botulinum neurotoxin X - high activity and unique substrate specificity
title Structural characterisation of the catalytic domain of botulinum neurotoxin X - high activity and unique substrate specificity
title_full Structural characterisation of the catalytic domain of botulinum neurotoxin X - high activity and unique substrate specificity
title_fullStr Structural characterisation of the catalytic domain of botulinum neurotoxin X - high activity and unique substrate specificity
title_full_unstemmed Structural characterisation of the catalytic domain of botulinum neurotoxin X - high activity and unique substrate specificity
title_short Structural characterisation of the catalytic domain of botulinum neurotoxin X - high activity and unique substrate specificity
title_sort structural characterisation of the catalytic domain of botulinum neurotoxin x - high activity and unique substrate specificity
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5851995/
https://www.ncbi.nlm.nih.gov/pubmed/29540745
http://dx.doi.org/10.1038/s41598-018-22842-4
work_keys_str_mv AT masuyergeoffrey structuralcharacterisationofthecatalyticdomainofbotulinumneurotoxinxhighactivityanduniquesubstratespecificity
AT zhangsicai structuralcharacterisationofthecatalyticdomainofbotulinumneurotoxinxhighactivityanduniquesubstratespecificity
AT barkhosulyman structuralcharacterisationofthecatalyticdomainofbotulinumneurotoxinxhighactivityanduniquesubstratespecificity
AT shenyi structuralcharacterisationofthecatalyticdomainofbotulinumneurotoxinxhighactivityanduniquesubstratespecificity
AT henrikssonlinda structuralcharacterisationofthecatalyticdomainofbotulinumneurotoxinxhighactivityanduniquesubstratespecificity
AT koseninasara structuralcharacterisationofthecatalyticdomainofbotulinumneurotoxinxhighactivityanduniquesubstratespecificity
AT dongmin structuralcharacterisationofthecatalyticdomainofbotulinumneurotoxinxhighactivityanduniquesubstratespecificity
AT stenmarkpal structuralcharacterisationofthecatalyticdomainofbotulinumneurotoxinxhighactivityanduniquesubstratespecificity