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Structure and conformational dynamics of Clostridioides difficile toxin A
Clostridioides difficile toxin A and B (TcdA and TcdB) are two major virulence factors responsible for diseases associated with C. difficile infection (CDI). Here, we report the 3.18-Å resolution crystal structure of a TcdA fragment (residues L843–T2481), which advances our understanding of the comp...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Life Science Alliance LLC
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8924006/ https://www.ncbi.nlm.nih.gov/pubmed/35292538 http://dx.doi.org/10.26508/lsa.202201383 |
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author | Chen, Baohua Basak, Sujit Chen, Peng Zhang, Changcheng Perry, Kay Tian, Songhai Yu, Clinton Dong, Min Huang, Lan Bowen, Mark E Jin, Rongsheng |
author_facet | Chen, Baohua Basak, Sujit Chen, Peng Zhang, Changcheng Perry, Kay Tian, Songhai Yu, Clinton Dong, Min Huang, Lan Bowen, Mark E Jin, Rongsheng |
author_sort | Chen, Baohua |
collection | PubMed |
description | Clostridioides difficile toxin A and B (TcdA and TcdB) are two major virulence factors responsible for diseases associated with C. difficile infection (CDI). Here, we report the 3.18-Å resolution crystal structure of a TcdA fragment (residues L843–T2481), which advances our understanding of the complete structure of TcdA holotoxin. Our structural analysis, together with complementary single molecule FRET and limited proteolysis studies, reveal that TcdA adopts a dynamic structure and its CROPs domain can sample a spectrum of open and closed conformations in a pH-dependent manner. Furthermore, a small globular subdomain (SGS) and the CROPs protect the pore-forming region of TcdA in the closed state at neutral pH, which could contribute to modulating the pH-dependent pore formation of TcdA. A rationally designed TcdA mutation that trapped the CROPs in the closed conformation showed drastically reduced cytotoxicity. Taken together, these studies shed new lights into the conformational dynamics of TcdA and its roles in TcdA intoxication. |
format | Online Article Text |
id | pubmed-8924006 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Life Science Alliance LLC |
record_format | MEDLINE/PubMed |
spelling | pubmed-89240062022-03-28 Structure and conformational dynamics of Clostridioides difficile toxin A Chen, Baohua Basak, Sujit Chen, Peng Zhang, Changcheng Perry, Kay Tian, Songhai Yu, Clinton Dong, Min Huang, Lan Bowen, Mark E Jin, Rongsheng Life Sci Alliance Research Articles Clostridioides difficile toxin A and B (TcdA and TcdB) are two major virulence factors responsible for diseases associated with C. difficile infection (CDI). Here, we report the 3.18-Å resolution crystal structure of a TcdA fragment (residues L843–T2481), which advances our understanding of the complete structure of TcdA holotoxin. Our structural analysis, together with complementary single molecule FRET and limited proteolysis studies, reveal that TcdA adopts a dynamic structure and its CROPs domain can sample a spectrum of open and closed conformations in a pH-dependent manner. Furthermore, a small globular subdomain (SGS) and the CROPs protect the pore-forming region of TcdA in the closed state at neutral pH, which could contribute to modulating the pH-dependent pore formation of TcdA. A rationally designed TcdA mutation that trapped the CROPs in the closed conformation showed drastically reduced cytotoxicity. Taken together, these studies shed new lights into the conformational dynamics of TcdA and its roles in TcdA intoxication. Life Science Alliance LLC 2022-03-15 /pmc/articles/PMC8924006/ /pubmed/35292538 http://dx.doi.org/10.26508/lsa.202201383 Text en © 2022 Chen et al. https://creativecommons.org/licenses/by/4.0/This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Research Articles Chen, Baohua Basak, Sujit Chen, Peng Zhang, Changcheng Perry, Kay Tian, Songhai Yu, Clinton Dong, Min Huang, Lan Bowen, Mark E Jin, Rongsheng Structure and conformational dynamics of Clostridioides difficile toxin A |
title | Structure and conformational dynamics of Clostridioides difficile toxin A |
title_full | Structure and conformational dynamics of Clostridioides difficile toxin A |
title_fullStr | Structure and conformational dynamics of Clostridioides difficile toxin A |
title_full_unstemmed | Structure and conformational dynamics of Clostridioides difficile toxin A |
title_short | Structure and conformational dynamics of Clostridioides difficile toxin A |
title_sort | structure and conformational dynamics of clostridioides difficile toxin a |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8924006/ https://www.ncbi.nlm.nih.gov/pubmed/35292538 http://dx.doi.org/10.26508/lsa.202201383 |
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