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Engineered MATE multidrug transporters reveal two functionally distinct ion-coupling pathways in NorM from Vibrio cholerae

Multidrug and toxic compound extrusion (MATE) transport proteins confer multidrug resistance on pathogenic microorganisms and affect pharmacokinetics in mammals. Our understanding of how MATE transporters work, has mostly relied on protein structures and MD simulations. However, the energetics of dr...

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Autores principales: Raturi, Sagar, Nair, Asha V., Shinoda, Keiko, Singh, Himansha, Bai, Boyan, Murakami, Satoshi, Fujitani, Hideaki, van Veen, Hendrik W.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8113278/
https://www.ncbi.nlm.nih.gov/pubmed/33976372
http://dx.doi.org/10.1038/s42003-021-02081-6
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author Raturi, Sagar
Nair, Asha V.
Shinoda, Keiko
Singh, Himansha
Bai, Boyan
Murakami, Satoshi
Fujitani, Hideaki
van Veen, Hendrik W.
author_facet Raturi, Sagar
Nair, Asha V.
Shinoda, Keiko
Singh, Himansha
Bai, Boyan
Murakami, Satoshi
Fujitani, Hideaki
van Veen, Hendrik W.
author_sort Raturi, Sagar
collection PubMed
description Multidrug and toxic compound extrusion (MATE) transport proteins confer multidrug resistance on pathogenic microorganisms and affect pharmacokinetics in mammals. Our understanding of how MATE transporters work, has mostly relied on protein structures and MD simulations. However, the energetics of drug transport has not been studied in detail. Many MATE transporters utilise the electrochemical H(+) or Na(+) gradient to drive substrate efflux, but NorM-VC from Vibrio cholerae can utilise both forms of metabolic energy. To dissect the localisation and organisation of H(+) and Na(+) translocation pathways in NorM-VC we engineered chimaeric proteins in which the N-lobe of H(+)-coupled NorM-PS from Pseudomonas stutzeri is fused to the C-lobe of NorM-VC, and vice versa. Our findings in drug binding and transport experiments with chimaeric, mutant and wildtype transporters highlight the versatile nature of energy coupling in NorM-VC, which enables adaptation to fluctuating salinity levels in the natural habitat of V. cholerae.
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spelling pubmed-81132782021-05-12 Engineered MATE multidrug transporters reveal two functionally distinct ion-coupling pathways in NorM from Vibrio cholerae Raturi, Sagar Nair, Asha V. Shinoda, Keiko Singh, Himansha Bai, Boyan Murakami, Satoshi Fujitani, Hideaki van Veen, Hendrik W. Commun Biol Article Multidrug and toxic compound extrusion (MATE) transport proteins confer multidrug resistance on pathogenic microorganisms and affect pharmacokinetics in mammals. Our understanding of how MATE transporters work, has mostly relied on protein structures and MD simulations. However, the energetics of drug transport has not been studied in detail. Many MATE transporters utilise the electrochemical H(+) or Na(+) gradient to drive substrate efflux, but NorM-VC from Vibrio cholerae can utilise both forms of metabolic energy. To dissect the localisation and organisation of H(+) and Na(+) translocation pathways in NorM-VC we engineered chimaeric proteins in which the N-lobe of H(+)-coupled NorM-PS from Pseudomonas stutzeri is fused to the C-lobe of NorM-VC, and vice versa. Our findings in drug binding and transport experiments with chimaeric, mutant and wildtype transporters highlight the versatile nature of energy coupling in NorM-VC, which enables adaptation to fluctuating salinity levels in the natural habitat of V. cholerae. Nature Publishing Group UK 2021-05-11 /pmc/articles/PMC8113278/ /pubmed/33976372 http://dx.doi.org/10.1038/s42003-021-02081-6 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/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/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Raturi, Sagar
Nair, Asha V.
Shinoda, Keiko
Singh, Himansha
Bai, Boyan
Murakami, Satoshi
Fujitani, Hideaki
van Veen, Hendrik W.
Engineered MATE multidrug transporters reveal two functionally distinct ion-coupling pathways in NorM from Vibrio cholerae
title Engineered MATE multidrug transporters reveal two functionally distinct ion-coupling pathways in NorM from Vibrio cholerae
title_full Engineered MATE multidrug transporters reveal two functionally distinct ion-coupling pathways in NorM from Vibrio cholerae
title_fullStr Engineered MATE multidrug transporters reveal two functionally distinct ion-coupling pathways in NorM from Vibrio cholerae
title_full_unstemmed Engineered MATE multidrug transporters reveal two functionally distinct ion-coupling pathways in NorM from Vibrio cholerae
title_short Engineered MATE multidrug transporters reveal two functionally distinct ion-coupling pathways in NorM from Vibrio cholerae
title_sort engineered mate multidrug transporters reveal two functionally distinct ion-coupling pathways in norm from vibrio cholerae
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8113278/
https://www.ncbi.nlm.nih.gov/pubmed/33976372
http://dx.doi.org/10.1038/s42003-021-02081-6
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