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Beltless Translocation Domain of Botulinum Neurotoxin A Embodies a Minimum Ion-conductive Channel
Botulinum neurotoxin, the causative agent of the paralytic disease botulism, is an endopeptidase composed of a catalytic domain (or light chain (LC)) and a heavy chain (HC) encompassing the translocation domain (TD) and receptor-binding domain. Upon receptor-mediated endocytosis, the LC and TD are p...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Biochemistry and Molecular Biology
2012
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3265847/ https://www.ncbi.nlm.nih.gov/pubmed/22158863 http://dx.doi.org/10.1074/jbc.C111.319400 |
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author | Fischer, Audrey Sambashivan, Shilpa Brunger, Axel T. Montal, Mauricio |
author_facet | Fischer, Audrey Sambashivan, Shilpa Brunger, Axel T. Montal, Mauricio |
author_sort | Fischer, Audrey |
collection | PubMed |
description | Botulinum neurotoxin, the causative agent of the paralytic disease botulism, is an endopeptidase composed of a catalytic domain (or light chain (LC)) and a heavy chain (HC) encompassing the translocation domain (TD) and receptor-binding domain. Upon receptor-mediated endocytosis, the LC and TD are proposed to undergo conformational changes in the acidic endocytic environment resulting in the formation of an LC protein-conducting TD channel. The mechanism of channel formation and the conformational changes in the toxin upon acidification are important but less well understood aspects of botulinum neurotoxin intoxication. Here, we have identified a minimum channel-forming truncation of the TD, the “beltless” TD, that forms transmembrane channels with ion conduction properties similar to those of the full-length TD. At variance with the holotoxin and the HC, channel formation for both the TD and the beltless TD occurs independent of a transmembrane pH gradient. Furthermore, acidification in solution induces moderate secondary structure changes. The subtle nature of the conformational changes evoked by acidification on the TD suggests that, in the context of the holotoxin, larger structural rearrangements and LC unfolding occur preceding or concurrent to channel formation. This notion is consistent with the hypothesis that although each domain of the holotoxin functions individually, each domain serves as a chaperone for the others. |
format | Online Article Text |
id | pubmed-3265847 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-32658472012-01-27 Beltless Translocation Domain of Botulinum Neurotoxin A Embodies a Minimum Ion-conductive Channel Fischer, Audrey Sambashivan, Shilpa Brunger, Axel T. Montal, Mauricio J Biol Chem Reports Botulinum neurotoxin, the causative agent of the paralytic disease botulism, is an endopeptidase composed of a catalytic domain (or light chain (LC)) and a heavy chain (HC) encompassing the translocation domain (TD) and receptor-binding domain. Upon receptor-mediated endocytosis, the LC and TD are proposed to undergo conformational changes in the acidic endocytic environment resulting in the formation of an LC protein-conducting TD channel. The mechanism of channel formation and the conformational changes in the toxin upon acidification are important but less well understood aspects of botulinum neurotoxin intoxication. Here, we have identified a minimum channel-forming truncation of the TD, the “beltless” TD, that forms transmembrane channels with ion conduction properties similar to those of the full-length TD. At variance with the holotoxin and the HC, channel formation for both the TD and the beltless TD occurs independent of a transmembrane pH gradient. Furthermore, acidification in solution induces moderate secondary structure changes. The subtle nature of the conformational changes evoked by acidification on the TD suggests that, in the context of the holotoxin, larger structural rearrangements and LC unfolding occur preceding or concurrent to channel formation. This notion is consistent with the hypothesis that although each domain of the holotoxin functions individually, each domain serves as a chaperone for the others. American Society for Biochemistry and Molecular Biology 2012-01-13 2011-12-12 /pmc/articles/PMC3265847/ /pubmed/22158863 http://dx.doi.org/10.1074/jbc.C111.319400 Text en © 2012 by The American Society for Biochemistry and Molecular Biology, Inc. Author's Choice—Final version full access. Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) applies to Author Choice Articles |
spellingShingle | Reports Fischer, Audrey Sambashivan, Shilpa Brunger, Axel T. Montal, Mauricio Beltless Translocation Domain of Botulinum Neurotoxin A Embodies a Minimum Ion-conductive Channel |
title | Beltless Translocation Domain of Botulinum Neurotoxin A Embodies a Minimum Ion-conductive Channel |
title_full | Beltless Translocation Domain of Botulinum Neurotoxin A Embodies a Minimum Ion-conductive Channel |
title_fullStr | Beltless Translocation Domain of Botulinum Neurotoxin A Embodies a Minimum Ion-conductive Channel |
title_full_unstemmed | Beltless Translocation Domain of Botulinum Neurotoxin A Embodies a Minimum Ion-conductive Channel |
title_short | Beltless Translocation Domain of Botulinum Neurotoxin A Embodies a Minimum Ion-conductive Channel |
title_sort | beltless translocation domain of botulinum neurotoxin a embodies a minimum ion-conductive channel |
topic | Reports |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3265847/ https://www.ncbi.nlm.nih.gov/pubmed/22158863 http://dx.doi.org/10.1074/jbc.C111.319400 |
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