Insight into the direct interaction of Na+ with NhaA and mechanistic implications

Abstract Na+/H+ antiporters comprise a family of membrane proteins evolutionarily conserved in all kingdoms of life that are essential in cellular ion homeostasis. While several human homologues have long been drug targets, NhaA of Escherichia coli has become the paradigm for this class of secondary...

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Autores principales:	Matthias Quick, Manish Dwivedi, Etana Padan
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/721bce5313a84b5289f7dd4b58c7b205
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spelling	oai:doaj.org-article:721bce5313a84b5289f7dd4b58c7b2052021-12-02T14:25:32ZInsight into the direct interaction of Na+ with NhaA and mechanistic implications10.1038/s41598-021-86318-82045-2322https://doaj.org/article/721bce5313a84b5289f7dd4b58c7b2052021-03-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-86318-8https://doaj.org/toc/2045-2322Abstract Na+/H+ antiporters comprise a family of membrane proteins evolutionarily conserved in all kingdoms of life that are essential in cellular ion homeostasis. While several human homologues have long been drug targets, NhaA of Escherichia coli has become the paradigm for this class of secondary active transporters as NhaA crystals provided insight in the structure of this molecular machine. However, structural data revealing the composition of the binding site for Na+ (or its surrogate Li+) is missing, representing a bottleneck in our understanding of the correlation between the structure and function of NhaA. Here, by adapting the scintillation proximity assay (SPA) for direct determination of Na+ binding to NhaA, we revealed that (i) NhaA is well adapted as the main antiporter for Na+ homeostasis in Escherichia coli and possibly in other bacteria as the cytoplasmic Na+ concentration is similar to the Na+ binding affinity of NhaA, (ii) experimental conditions affect NhaA-mediated cation binding, (iii) in addition to Na+ and Li+, the halide Tl+ interacts with NhaA, (iv) whereas acidic pH inhibits maximum binding of Na+ to NhaA, partial Na+ binding by NhaA is independent of the pH, an important novel insight into the effect of pH on NhaA cation binding.Matthias QuickManish DwivediEtana PadanNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-9 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Matthias Quick Manish Dwivedi Etana Padan Insight into the direct interaction of Na+ with NhaA and mechanistic implications
description	Abstract Na+/H+ antiporters comprise a family of membrane proteins evolutionarily conserved in all kingdoms of life that are essential in cellular ion homeostasis. While several human homologues have long been drug targets, NhaA of Escherichia coli has become the paradigm for this class of secondary active transporters as NhaA crystals provided insight in the structure of this molecular machine. However, structural data revealing the composition of the binding site for Na+ (or its surrogate Li+) is missing, representing a bottleneck in our understanding of the correlation between the structure and function of NhaA. Here, by adapting the scintillation proximity assay (SPA) for direct determination of Na+ binding to NhaA, we revealed that (i) NhaA is well adapted as the main antiporter for Na+ homeostasis in Escherichia coli and possibly in other bacteria as the cytoplasmic Na+ concentration is similar to the Na+ binding affinity of NhaA, (ii) experimental conditions affect NhaA-mediated cation binding, (iii) in addition to Na+ and Li+, the halide Tl+ interacts with NhaA, (iv) whereas acidic pH inhibits maximum binding of Na+ to NhaA, partial Na+ binding by NhaA is independent of the pH, an important novel insight into the effect of pH on NhaA cation binding.
format	article
author	Matthias Quick Manish Dwivedi Etana Padan
author_facet	Matthias Quick Manish Dwivedi Etana Padan
author_sort	Matthias Quick
title	Insight into the direct interaction of Na+ with NhaA and mechanistic implications
title_short	Insight into the direct interaction of Na+ with NhaA and mechanistic implications
title_full	Insight into the direct interaction of Na+ with NhaA and mechanistic implications
title_fullStr	Insight into the direct interaction of Na+ with NhaA and mechanistic implications
title_full_unstemmed	Insight into the direct interaction of Na+ with NhaA and mechanistic implications
title_sort	insight into the direct interaction of na+ with nhaa and mechanistic implications
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/721bce5313a84b5289f7dd4b58c7b205
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Insight into the direct interaction of Na+ with NhaA and mechanistic implications

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