Crystal structure of the sodium-proton antiporter NhaA dimer and new mechanistic insights

Chiara Lee, Shoko Yashiro, David L. Dotson, Povilas Uzdavinys, So Iwata, Mark S P Sansom, Christoph von Ballmoos, Oliver Beckstein, David Drew, Alexander D. Cameron

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Sodium-proton antiporters rapidly exchange protons and sodium ions across the membrane to regulate intracellular pH, cell volume, and sodium concentration. How ion binding and release is coupled to the conformational changes associated with transport is not clear. Here, we report a crystal form of the prototypical sodium-proton antiporter NhaA from Escherichia coli in which the protein is seen as a dimer. In this new structure, we observe a salt bridge between an essential aspartic acid (Asp163) and a conserved lysine (Lys300). An equivalent salt bridge is present in the homologous transporter NapA, but not in the only other known crystal structure of NhaA, which provides the foundation of most existing structural models of electrogenic sodium-proton antiport. Molecular dynamics simulations show that the stability of the salt bridge is weakened by sodium ions binding to Asp164 and the neighboring Asp163. This suggests that the transport mechanism involves Asp163 switching between forming a salt bridge with Lys300 and interacting with the sodium ion. pKa calculations suggest that Asp163 is highly unlikely to be protonated when involved in the salt bridge. As it has been previously suggested that Asp163 is one of the two residues through which proton transport occurs, these results have clear implications to the current mechanistic models of sodium-proton antiport in NhaA.

Original languageEnglish (US)
Pages (from-to)529-544
Number of pages16
JournalJournal of General Physiology
Volume144
Issue number6
DOIs
StatePublished - 2014

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Sodium-Hydrogen Antiporter
Sodium
Salts
Protons
Ions
Ion Transport
Structural Models
Molecular Dynamics Simulation
Cell Size
Aspartic Acid
Lysine
Escherichia coli
Membranes
Proteins

ASJC Scopus subject areas

  • Physiology

Cite this

Lee, C., Yashiro, S., Dotson, D. L., Uzdavinys, P., Iwata, S., Sansom, M. S. P., ... Cameron, A. D. (2014). Crystal structure of the sodium-proton antiporter NhaA dimer and new mechanistic insights. Journal of General Physiology, 144(6), 529-544. https://doi.org/10.1085/jgp.201411219

Crystal structure of the sodium-proton antiporter NhaA dimer and new mechanistic insights. / Lee, Chiara; Yashiro, Shoko; Dotson, David L.; Uzdavinys, Povilas; Iwata, So; Sansom, Mark S P; von Ballmoos, Christoph; Beckstein, Oliver; Drew, David; Cameron, Alexander D.

In: Journal of General Physiology, Vol. 144, No. 6, 2014, p. 529-544.

Research output: Contribution to journalArticle

Lee, C, Yashiro, S, Dotson, DL, Uzdavinys, P, Iwata, S, Sansom, MSP, von Ballmoos, C, Beckstein, O, Drew, D & Cameron, AD 2014, 'Crystal structure of the sodium-proton antiporter NhaA dimer and new mechanistic insights', Journal of General Physiology, vol. 144, no. 6, pp. 529-544. https://doi.org/10.1085/jgp.201411219
Lee, Chiara ; Yashiro, Shoko ; Dotson, David L. ; Uzdavinys, Povilas ; Iwata, So ; Sansom, Mark S P ; von Ballmoos, Christoph ; Beckstein, Oliver ; Drew, David ; Cameron, Alexander D. / Crystal structure of the sodium-proton antiporter NhaA dimer and new mechanistic insights. In: Journal of General Physiology. 2014 ; Vol. 144, No. 6. pp. 529-544.
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