title: A Single-Pore Residue Renders the Arabidopsis Root Anion Channel SLAH2 Highly Nitrate Selective.
abstract: In contrast to animal cells, plants use nitrate as a major source of nitrogen. Following the uptake of nitrate, this major macronutrient is fed into the vasculature for long-distance transport. The Arabidopsis thaliana shoot expresses the anion channel SLOW ANION CHANNEL1 (SLAC1) and its homolog SLAC1 HOMOLOGOUS3 (SLAH3), which prefer nitrate as substrate but cannot exclude chloride ions. By contrast, we identified SLAH2 as a nitrate-specific channel that is impermeable for chloride. To understand the molecular basis for nitrate selection in the SLAH2 channel, SLAC1 and SLAH2 were modeled to the structure of HiTehA, a distantly related bacterial member. Structure-guided site-directed mutations converted SLAC1 into a SLAH2-like nitrate-specific anion channel and vice versa. Our findings indicate that two pore-occluding phenylalanines constrict the pore. The selectivity filter of SLAC/SLAH anion channels is determined by the polarity of pore-lining residues located on alpha helix 3. Changing the polar character of a single amino acid side chain (Ser-228) to a nonpolar residue turned the nitrate-selective SLAH2 into a chloride/nitrate-permeable anion channel. Thus, the molecular basis of the anion specificity of SLAC/SLAH anion channels seems to be determined by the presence and constellation of polar side chains that act in concert with the two pore-occluding phenylalanines.
authors.affiliation: University of Würzburg, Institute for Molecular Plant Physiology and Biophysics, D-97082 Würzburg, Germany.
authors.forename: Tobias
authors.initials: T
authors.lastname: Maierhofer
authors.affiliation: University of Würzburg, Institute for Molecular Plant Physiology and Biophysics, D-97082 Würzburg, Germany.
authors.forename: Christof
authors.initials: C
authors.lastname: Lind
authors.affiliation: University of Würzburg, Institute for Molecular Plant Physiology and Biophysics, D-97082 Würzburg, Germany.
authors.forename: Stefanie
authors.initials: S
authors.lastname: Hüttl
authors.affiliation: University of Würzburg, Institute for Molecular Plant Physiology and Biophysics, D-97082 Würzburg, Germany.
authors.forename: Sönke
authors.initials: S
authors.lastname: Scherzer
authors.affiliation: University of Würzburg, Institute for Molecular Plant Physiology and Biophysics, D-97082 Würzburg, Germany.
authors.forename: Melanie
authors.initials: M
authors.lastname: Papenfuß
authors.affiliation: Institute of Forest Sciences, Chair of Tree Physiology, University of Freiburg, 79110 Freiburg, Germany.
authors.forename: Judy
authors.initials: J
authors.lastname: Simon
authors.affiliation: Zoology Department, College of Science, King Saud University, Riyadh 11451, Saudi Arabia.
authors.forename: Khaled A S
authors.initials: KA
authors.lastname: Al-Rasheid
authors.affiliation: University of Würzburg, Institute for Molecular Plant Physiology and Biophysics, D-97082 Würzburg, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Ache
authors.affiliation: Institute of Forest Sciences, Chair of Tree Physiology, University of Freiburg, 79110 Freiburg, Germany.
authors.forename: Heinz
authors.initials: H
authors.lastname: Rennenberg
authors.affiliation: University of Würzburg, Institute for Molecular Plant Physiology and Biophysics, D-97082 Würzburg, Germany.
authors.forename: Rainer
authors.initials: R
authors.lastname: Hedrich
authors.affiliation: University of Würzburg, Institute for Molecular Plant Physiology and Biophysics, D-97082 Würzburg, Germany.
authors.forename: Thomas D
authors.initials: TD
authors.lastname: Müller
authors.affiliation: University of Würzburg, Institute for Molecular Plant Physiology and Biophysics, D-97082 Würzburg, Germany geiger@botanik.uni-wuerzburg.de.
authors.forename: Dietmar
authors.identifier: http://orcid.org/0000-0003-0715-5710
authors.initials: D
authors.lastname: Geiger