Protein architecture and core residues in unwound α-helices provide insights to the transport function of plant AtCHX17

Czerny, Daniel D.; Padmanaban, Senthilkumar; Anishkin, Andriy; Venema, Kees; Riaz, Zoya; Sze, Heven

doi:10.1016/j.bbamem.2016.05.008

Title: Protein architecture and core residues in unwound α-helices provide insights to the transport function of plant AtCHX17

Journal Article · Thu Sep 01 00:00:00 EDT 2016 · Biochimica et Biophysica Acta. Biomembranes

DOI:https://doi.org/10.1016/j.bbamem.2016.05.008· OSTI ID:1325359

Czerny, Daniel D.; Padmanaban, Senthilkumar; Anishkin, Andriy; Venema, Kees; Riaz, Zoya; Sze, Heven

Using Arabidopsis thaliana AtCHX17 as an example, we combine structural modeling and mutagenesis to provide insights on its protein architecture and transport function which is poorly characterized. This approach is based on the observation that protein structures are significantly more conserved in evolution than linear sequences, and mechanistic similarities among diverse transporters are emerging. Two homology models of AtCHX17 were obtained that show a protein fold similar to known structures of bacterial Na⁺/H⁺ antiporters, EcNhaA and TtNapA. The distinct secondary and tertiary structure models highlighted residues at positions potentially important for CHX17 activity. Mutagenesis showed that asparagine-N200 and aspartate-D201 inside transmembrane5 (TM5), and lysine-K355 inside TM10 are critical for AtCHX17 activity. We reveal previously unrecognized threonine-T170 and lysine-K383 as key residues at unwound regions in the middle of TM4 and TM11 α-helices, respectively. Mutation of glutamate-E111 located near the membrane surface inhibited AtCHX17 activity, suggesting a role in pH sensing. The long carboxylic tail of unknown purpose has an alternating β-sheet and α-helix secondary structure that is conserved in prokaryote universal stress proteins. Here, these results support the overall architecture of AtCHX17 and identify D201, N200 and novel residues T170 and K383 at the functional core which likely participates in ion recognition, coordination and/or translocation, similar to characterized cation/H⁺ exchangers. The core of AtCHX17 models according to EcNhaA and TtNapA templates faces inward and outward, respectively, which may reflect two conformational states of the alternating access transport mode for proteins belonging to the plant CHX family.

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Research Organization:: Univ. of Maryland, College Park, MD (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES)

Contributing Organization:: University of Maryland

Grant/Contract Number:: BES DEFG-0207ER15883; BIO2012-33655; BIO2015-65056; FG02-07ER15883

OSTI ID:: 1325359

Alternate ID(s):: OSTI ID: 1351089

Journal Information:: Biochimica et Biophysica Acta. Biomembranes, Journal Name: Biochimica et Biophysica Acta. Biomembranes Vol. 1858 Journal Issue: 9; ISSN 0005-2736

Publisher:: ElsevierCopyright Statement

Country of Publication:: Netherlands

Language:: English

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Cited by: 14 works

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Title: Protein architecture and core residues in unwound α-helices provide insights to the transport function of plant AtCHX17

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