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Deciphering ion transport and ATPase coupling in the intersubunit tunnel of KdpFABC

KdpFABC, a high-affinity K(+) pump, combines the ion channel KdpA and the P-type ATPase KdpB to secure survival at K(+) limitation. Here, we apply a combination of cryo-EM, biochemical assays, and MD simulations to illuminate the mechanisms underlying transport and the coupling to ATP hydrolysis. We...

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Autores principales: Silberberg, Jakob M., Corey, Robin A., Hielkema, Lisa, Stock, Charlott, Stansfeld, Phillip J., Paulino, Cristina, Hänelt, Inga
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8385062/
https://www.ncbi.nlm.nih.gov/pubmed/34429416
http://dx.doi.org/10.1038/s41467-021-25242-x
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author Silberberg, Jakob M.
Corey, Robin A.
Hielkema, Lisa
Stock, Charlott
Stansfeld, Phillip J.
Paulino, Cristina
Hänelt, Inga
author_facet Silberberg, Jakob M.
Corey, Robin A.
Hielkema, Lisa
Stock, Charlott
Stansfeld, Phillip J.
Paulino, Cristina
Hänelt, Inga
author_sort Silberberg, Jakob M.
collection PubMed
description KdpFABC, a high-affinity K(+) pump, combines the ion channel KdpA and the P-type ATPase KdpB to secure survival at K(+) limitation. Here, we apply a combination of cryo-EM, biochemical assays, and MD simulations to illuminate the mechanisms underlying transport and the coupling to ATP hydrolysis. We show that ions are transported via an intersubunit tunnel through KdpA and KdpB. At the subunit interface, the tunnel is constricted by a phenylalanine, which, by polarized cation-π stacking, controls K(+) entry into the canonical substrate binding site (CBS) of KdpB. Within the CBS, ATPase coupling is mediated by the charge distribution between an aspartate and a lysine. Interestingly, individual elements of the ion translocation mechanism of KdpFABC identified here are conserved among a wide variety of P-type ATPases from different families. This leads us to the hypothesis that KdpB might represent an early descendant of a common ancestor of cation pumps.
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spelling pubmed-83850622021-09-22 Deciphering ion transport and ATPase coupling in the intersubunit tunnel of KdpFABC Silberberg, Jakob M. Corey, Robin A. Hielkema, Lisa Stock, Charlott Stansfeld, Phillip J. Paulino, Cristina Hänelt, Inga Nat Commun Article KdpFABC, a high-affinity K(+) pump, combines the ion channel KdpA and the P-type ATPase KdpB to secure survival at K(+) limitation. Here, we apply a combination of cryo-EM, biochemical assays, and MD simulations to illuminate the mechanisms underlying transport and the coupling to ATP hydrolysis. We show that ions are transported via an intersubunit tunnel through KdpA and KdpB. At the subunit interface, the tunnel is constricted by a phenylalanine, which, by polarized cation-π stacking, controls K(+) entry into the canonical substrate binding site (CBS) of KdpB. Within the CBS, ATPase coupling is mediated by the charge distribution between an aspartate and a lysine. Interestingly, individual elements of the ion translocation mechanism of KdpFABC identified here are conserved among a wide variety of P-type ATPases from different families. This leads us to the hypothesis that KdpB might represent an early descendant of a common ancestor of cation pumps. Nature Publishing Group UK 2021-08-24 /pmc/articles/PMC8385062/ /pubmed/34429416 http://dx.doi.org/10.1038/s41467-021-25242-x 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
Silberberg, Jakob M.
Corey, Robin A.
Hielkema, Lisa
Stock, Charlott
Stansfeld, Phillip J.
Paulino, Cristina
Hänelt, Inga
Deciphering ion transport and ATPase coupling in the intersubunit tunnel of KdpFABC
title Deciphering ion transport and ATPase coupling in the intersubunit tunnel of KdpFABC
title_full Deciphering ion transport and ATPase coupling in the intersubunit tunnel of KdpFABC
title_fullStr Deciphering ion transport and ATPase coupling in the intersubunit tunnel of KdpFABC
title_full_unstemmed Deciphering ion transport and ATPase coupling in the intersubunit tunnel of KdpFABC
title_short Deciphering ion transport and ATPase coupling in the intersubunit tunnel of KdpFABC
title_sort deciphering ion transport and atpase coupling in the intersubunit tunnel of kdpfabc
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8385062/
https://www.ncbi.nlm.nih.gov/pubmed/34429416
http://dx.doi.org/10.1038/s41467-021-25242-x
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