Numerous essential physiological processes depend on the TMEM16A-mediated Ca2+-activated chloride fluxes. Extensive structure–function studies have helped to elucidate the Ca2+ gating mechanism of TMEM16A, revealing a Ca2+-sensing element close to the anion pore that alters conduction. However, substrate selection and the substrate–gating relationship in TMEM16A remain less explored. Here, we study the gating–permeant anion relationship on mouse TMEM16A expressed in HEK 293 cells using electrophysiological recordings coupled with site-directed mutagenesis. We show that the apparent Ca2+ sensitivity of TMEM16A increased with highly permeant anions and SCN− mole fractions, likely by stabilizing bound Ca2+. Conversely, mutations at crucial gating elements, including the Ca2+-binding site 1, the transmembrane helix 6 (TM6), and the hydrophobic gate, impaired the anion permeability and selectivity of TMEM16A. Finally, we found that, unlike anion-selective wild-type channels, the voltage dependence of unselective TMEM16A mutant channels was less sensitive to SCN−. Therefore, our work identifies structural determinants of selectivity at the Ca2+ site, TM6, and hydrophobic gate and reveals a reciprocal regulation of gating and selectivity. We suggest that this regulation is essential to set ionic selectivity and the Ca2+ and voltage sensitivities in TMEM16A.
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Communication|
June 10 2022
Gating and anion selectivity are reciprocally regulated in TMEM16A (ANO1)
José J. De Jesús-Pérez
,
José J. De Jesús-Pérez
1
Physics Institute, Universidad Autónoma de San Luis Potosí, San Luis Potosí, México
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Ana E. López-Romero
,
Ana E. López-Romero
1
Physics Institute, Universidad Autónoma de San Luis Potosí, San Luis Potosí, México
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Odalys Posadas,
Odalys Posadas
1
Physics Institute, Universidad Autónoma de San Luis Potosí, San Luis Potosí, México
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Guadalupe Segura-Covarrubias
,
Guadalupe Segura-Covarrubias
1
Physics Institute, Universidad Autónoma de San Luis Potosí, San Luis Potosí, México
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Iván Aréchiga-Figueroa,
Iván Aréchiga-Figueroa
2
Consejo Nacional de Ciencia y Tecnología, Facultad de Medicina, Universidad Autónoma de San Luis Potosí, San Luis Potosí, México
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Braulio Gutiérrez-Medina,
Braulio Gutiérrez-Medina
3
Advanced Materials Division, Instituto Potosino de Investigación Científica y Tecnológica, San Luis Potosí, México
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Patricia Pérez-Cornejo
,
Patricia Pérez-Cornejo
4
Department of Physiology and Biophysics, Facultad de Medicina, Universidad Autónoma de San Luis Potosí, San Luis Potosí, México
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Jorge Arreola
1
Physics Institute, Universidad Autónoma de San Luis Potosí, San Luis Potosí, México
Correspondence to Jorge Arreola: arreola@dec1.ifisica.uaslp.mx
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José J. De Jesús-Pérez
1
Physics Institute, Universidad Autónoma de San Luis Potosí, San Luis Potosí, México
Ana E. López-Romero
1
Physics Institute, Universidad Autónoma de San Luis Potosí, San Luis Potosí, México
Odalys Posadas
1
Physics Institute, Universidad Autónoma de San Luis Potosí, San Luis Potosí, México
Guadalupe Segura-Covarrubias
1
Physics Institute, Universidad Autónoma de San Luis Potosí, San Luis Potosí, México
Iván Aréchiga-Figueroa
2
Consejo Nacional de Ciencia y Tecnología, Facultad de Medicina, Universidad Autónoma de San Luis Potosí, San Luis Potosí, México
Braulio Gutiérrez-Medina
3
Advanced Materials Division, Instituto Potosino de Investigación Científica y Tecnológica, San Luis Potosí, México
Patricia Pérez-Cornejo
4
Department of Physiology and Biophysics, Facultad de Medicina, Universidad Autónoma de San Luis Potosí, San Luis Potosí, México
Correspondence to Jorge Arreola: arreola@dec1.ifisica.uaslp.mx
J.J. De Jesús-Pérez’s current address is Department of Systems Pharmacology and Translational Therapeutics, University of Pennsylvania, Perelman School of Medicine, Philadelphia, PA.
G. Segura-Covarrubias’s current address is Department of Physiology and Biophysics, Case Western Reserve University, Cleveland, OH.
Received:
September 17 2021
Accepted:
May 23 2022
Online Issn: 1540-7748
Print Issn: 0022-1295
Funding
Funder(s):
FORDECYT-PRONACES
- Award Id(s): 1308052 2020
Funder(s):
Consejo Nacional de Ciencia y Tecnología
- Award Id(s): 234820,711128,933084,297721
© 2022 De Jesús-Pérez et al.
2022
This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/).
J Gen Physiol (2022) 154 (8): e202113027.
Article history
Received:
September 17 2021
Accepted:
May 23 2022
Citation
José J. De Jesús-Pérez, Ana E. López-Romero, Odalys Posadas, Guadalupe Segura-Covarrubias, Iván Aréchiga-Figueroa, Braulio Gutiérrez-Medina, Patricia Pérez-Cornejo, Jorge Arreola; Gating and anion selectivity are reciprocally regulated in TMEM16A (ANO1). J Gen Physiol 1 August 2022; 154 (8): e202113027. doi: https://doi.org/10.1085/jgp.202113027
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