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Formation of Fibrils by the Periplasmic Molecular Chaperone HdeB from Escherichia coli

The molecular chaperones HdeA and HdeB of the Escherichia coli (E. coli) periplasm protect client proteins from acid denaturation through a unique mechanism that utilizes their acid denatured states to bind clients. We previously demonstrated that the active, acid-denatured form of HdeA is also pron...

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Autores principales: Nakata, Yui, Kitazaki, Yuuto, Kanaoka, Hitomi, Shingen, Erika, Uehara, Rina, Hongo, Kunihiro, Kawata, Yasushi, Mizobata, Tomohiro
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9657021/
https://www.ncbi.nlm.nih.gov/pubmed/36362039
http://dx.doi.org/10.3390/ijms232113243
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author Nakata, Yui
Kitazaki, Yuuto
Kanaoka, Hitomi
Shingen, Erika
Uehara, Rina
Hongo, Kunihiro
Kawata, Yasushi
Mizobata, Tomohiro
author_facet Nakata, Yui
Kitazaki, Yuuto
Kanaoka, Hitomi
Shingen, Erika
Uehara, Rina
Hongo, Kunihiro
Kawata, Yasushi
Mizobata, Tomohiro
author_sort Nakata, Yui
collection PubMed
description The molecular chaperones HdeA and HdeB of the Escherichia coli (E. coli) periplasm protect client proteins from acid denaturation through a unique mechanism that utilizes their acid denatured states to bind clients. We previously demonstrated that the active, acid-denatured form of HdeA is also prone to forming inactive, amyloid fibril-like aggregates in a pH-dependent, reversible manner. In this study, we report that HdeB also displays a similar tendency to form fibrils at low pH. HdeB fibrils were observed at pH < 3 in the presence of NaCl. Similar to HdeA, HdeB fibrils could be resolubilized by a simple shift to neutral pH. In the case of HdeB, however, we found that after extended incubation at low pH, HdeB fibrils were converted into a form that could not resolubilize at pH 7. Fresh fibrils seeded from these “transformed” fibrils were also incapable of resolubilizing at pH 7, suggesting that the transition from reversible to irreversible fibrils involved a specific conformational change that was transmissible through fibril seeds. Analyses of fibril secondary structure indicated that HdeB fibrils retained significant alpha helical content regardless of the conditions under which fibrils were formed. Fibrils that were formed from HdeB that had been treated to remove its intrinsic disulfide bond also were incapable of resolubilizing at pH 7, suggesting that certain residual structures that are retained in acid-denatured HdeB are important for this protein to recover its soluble state from the fibril form.
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spelling pubmed-96570212022-11-15 Formation of Fibrils by the Periplasmic Molecular Chaperone HdeB from Escherichia coli Nakata, Yui Kitazaki, Yuuto Kanaoka, Hitomi Shingen, Erika Uehara, Rina Hongo, Kunihiro Kawata, Yasushi Mizobata, Tomohiro Int J Mol Sci Article The molecular chaperones HdeA and HdeB of the Escherichia coli (E. coli) periplasm protect client proteins from acid denaturation through a unique mechanism that utilizes their acid denatured states to bind clients. We previously demonstrated that the active, acid-denatured form of HdeA is also prone to forming inactive, amyloid fibril-like aggregates in a pH-dependent, reversible manner. In this study, we report that HdeB also displays a similar tendency to form fibrils at low pH. HdeB fibrils were observed at pH < 3 in the presence of NaCl. Similar to HdeA, HdeB fibrils could be resolubilized by a simple shift to neutral pH. In the case of HdeB, however, we found that after extended incubation at low pH, HdeB fibrils were converted into a form that could not resolubilize at pH 7. Fresh fibrils seeded from these “transformed” fibrils were also incapable of resolubilizing at pH 7, suggesting that the transition from reversible to irreversible fibrils involved a specific conformational change that was transmissible through fibril seeds. Analyses of fibril secondary structure indicated that HdeB fibrils retained significant alpha helical content regardless of the conditions under which fibrils were formed. Fibrils that were formed from HdeB that had been treated to remove its intrinsic disulfide bond also were incapable of resolubilizing at pH 7, suggesting that certain residual structures that are retained in acid-denatured HdeB are important for this protein to recover its soluble state from the fibril form. MDPI 2022-10-31 /pmc/articles/PMC9657021/ /pubmed/36362039 http://dx.doi.org/10.3390/ijms232113243 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Nakata, Yui
Kitazaki, Yuuto
Kanaoka, Hitomi
Shingen, Erika
Uehara, Rina
Hongo, Kunihiro
Kawata, Yasushi
Mizobata, Tomohiro
Formation of Fibrils by the Periplasmic Molecular Chaperone HdeB from Escherichia coli
title Formation of Fibrils by the Periplasmic Molecular Chaperone HdeB from Escherichia coli
title_full Formation of Fibrils by the Periplasmic Molecular Chaperone HdeB from Escherichia coli
title_fullStr Formation of Fibrils by the Periplasmic Molecular Chaperone HdeB from Escherichia coli
title_full_unstemmed Formation of Fibrils by the Periplasmic Molecular Chaperone HdeB from Escherichia coli
title_short Formation of Fibrils by the Periplasmic Molecular Chaperone HdeB from Escherichia coli
title_sort formation of fibrils by the periplasmic molecular chaperone hdeb from escherichia coli
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9657021/
https://www.ncbi.nlm.nih.gov/pubmed/36362039
http://dx.doi.org/10.3390/ijms232113243
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