Protein architecture and core residues in unwound α-helices provide insights to the transport function of plant AtCHX17
Abstract
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 inmore »
- Authors:
- Publication Date:
- Research Org.:
- Univ. of Maryland, College Park, MD (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Basic Energy Sciences (BES)
- Contributing Org.:
- University of Maryland
- OSTI Identifier:
- 1325359
- Alternate Identifier(s):
- OSTI ID: 1351089
- Grant/Contract Number:
- BES DEFG-0207ER15883; BIO2012-33655; BIO2015-65056; FG02-07ER15883
- Resource Type:
- Published Article
- Journal Name:
- Biochimica et Biophysica Acta. Biomembranes
- Additional Journal Information:
- Journal Name: Biochimica et Biophysica Acta. Biomembranes Journal Volume: 1858 Journal Issue: 9; Journal ID: ISSN 0005-2736
- Publisher:
- Elsevier
- Country of Publication:
- Netherlands
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; cation/proton antiporter; potassium; structure modeling; transporter; pH regulation; yeast expression; Arabidopsis; core residues
Citation Formats
Czerny, Daniel D., Padmanaban, Senthilkumar, Anishkin, Andriy, Venema, Kees, Riaz, Zoya, and Sze, Heven. Protein architecture and core residues in unwound α-helices provide insights to the transport function of plant AtCHX17. Netherlands: N. p., 2016.
Web. doi:10.1016/j.bbamem.2016.05.008.
Czerny, Daniel D., Padmanaban, Senthilkumar, Anishkin, Andriy, Venema, Kees, Riaz, Zoya, & Sze, Heven. Protein architecture and core residues in unwound α-helices provide insights to the transport function of plant AtCHX17. Netherlands. https://doi.org/10.1016/j.bbamem.2016.05.008
Czerny, Daniel D., Padmanaban, Senthilkumar, Anishkin, Andriy, Venema, Kees, Riaz, Zoya, and Sze, Heven. Thu .
"Protein architecture and core residues in unwound α-helices provide insights to the transport function of plant AtCHX17". Netherlands. https://doi.org/10.1016/j.bbamem.2016.05.008.
@article{osti_1325359,
title = {Protein architecture and core residues in unwound α-helices provide insights to the transport function of plant AtCHX17},
author = {Czerny, Daniel D. and Padmanaban, Senthilkumar and Anishkin, Andriy and Venema, Kees and Riaz, Zoya and Sze, Heven},
abstractNote = {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.},
doi = {10.1016/j.bbamem.2016.05.008},
journal = {Biochimica et Biophysica Acta. Biomembranes},
number = 9,
volume = 1858,
place = {Netherlands},
year = {Thu Sep 01 00:00:00 EDT 2016},
month = {Thu Sep 01 00:00:00 EDT 2016}
}
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