title: Conformations of NhaA, the Na/H exchanger from Escherichia coli, in the pH-activated and ion-translocating states.
abstract: NhaA, the main sodium-proton exchanger in the inner membrane of Escherichia coli, regulates the cytosolic concentrations of H and Na. It is inactive at acidic pH, becomes active between pH 6 and pH 7, and reaches maximum activity at pH 8. By cryo-electron microscopy of two-dimensional crystals grown at pH 4 and incubated at higher pH, we identified two sequential conformational changes in the protein in response to pH or substrate ions. The first change is induced by a rise in pH from 6 to 7 and marks the transition from the inactive state to the pH-activated state. pH activation, which precedes the ion-induced conformational change, is accompanied by an overall expansion of the NhaA monomer and a local ordering of the N-terminus. The second conformational change is induced by the substrate ions Na and Li at pH above 7 and involves a 7-A displacement of helix IVp. This movement would cause a charge imbalance at the ion-binding site that may trigger the release of the substrate ion and open a periplasmic exit channel.
authors.affiliation: Department of Structural Biology, Max Planck Institute of Biophysics, Frankfurt am Main, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Appel
authors.forename: Dilem
authors.initials: D
authors.lastname: Hizlan
authors.forename: Kutti R
authors.initials: KR
authors.lastname: Vinothkumar
authors.forename: Christine
authors.initials: C
authors.lastname: Ziegler
authors.forename: Werner
authors.initials: W
authors.lastname: Kühlbrandt
mesh_terms: D020285:Cryoelectron Microscopy; D004926:Escherichia coli / Q000502:physiology*; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000648:ultrastructure; D006863:Hydrogen-Ion Concentration; D008958:Models, Molecular; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D011522:Protons; D012964:Sodium; D017923:Sodium-Hydrogen Exchangers / Q000737:chemistry* / Q000648:ultrastructure
chemical_list: D029968:Escherichia coli Proteins; C102839:NhaA protein, E coli; D011522:Protons; D017923:Sodium-Hydrogen Exchangers; D012964:Sodium