Electro-steric opening of the clc-2 chloride channel gate

Abstract The widely expressed two-pore homodimeric inward rectifier CLC-2 chloride channel regulates transepithelial chloride transport, extracellular chloride homeostasis, and neuronal excitability. Each pore is independently gated at hyperpolarized voltages by a conserved pore glutamate. Presumabl...

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Autores principales:	José J. De Jesús-Pérez, G. Arlette Méndez-Maldonado, Ana E. López-Romero, David Esparza-Jasso, Irma L. González-Hernández, Víctor De la Rosa, Roberto Gastélum-Garibaldi, Jorge E. Sánchez-Rodríguez, Jorge Arreola
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/d68ca9a572f94156b3003b15ab5f24b7
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spelling	oai:doaj.org-article:d68ca9a572f94156b3003b15ab5f24b72021-12-02T16:06:10ZElectro-steric opening of the clc-2 chloride channel gate10.1038/s41598-021-92247-32045-2322https://doaj.org/article/d68ca9a572f94156b3003b15ab5f24b72021-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-92247-3https://doaj.org/toc/2045-2322Abstract The widely expressed two-pore homodimeric inward rectifier CLC-2 chloride channel regulates transepithelial chloride transport, extracellular chloride homeostasis, and neuronal excitability. Each pore is independently gated at hyperpolarized voltages by a conserved pore glutamate. Presumably, exiting chloride ions push glutamate outwardly while external protonation stabilizes it. To understand the mechanism of mouse CLC-2 opening we used homology modelling-guided structure–function analysis. Structural modelling suggests that glutamate E213 interacts with tyrosine Y561 to close a pore. Accordingly, Y561A and E213D mutants are activated at less hyperpolarized voltages, re-opened at depolarized voltages, and fast and common gating components are reduced. The double mutant cycle analysis showed that E213 and Y561 are energetically coupled to alter CLC-2 gating. In agreement, the anomalous mole fraction behaviour of the voltage dependence, measured by the voltage to induce half-open probability, was strongly altered in these mutants. Finally, cytosolic acidification or high extracellular chloride concentration, conditions that have little or no effect on WT CLC-2, induced reopening of Y561 mutants at positive voltages presumably by the inward opening of E213. We concluded that the CLC-2 gate is formed by Y561-E213 and that outward permeant anions open the gate by electrostatic and steric interactions.José J. De Jesús-PérezG. Arlette Méndez-MaldonadoAna E. López-RomeroDavid Esparza-JassoIrma L. González-HernándezVíctor De la RosaRoberto Gastélum-GaribaldiJorge E. Sánchez-RodríguezJorge ArreolaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-14 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q José J. De Jesús-Pérez G. Arlette Méndez-Maldonado Ana E. López-Romero David Esparza-Jasso Irma L. González-Hernández Víctor De la Rosa Roberto Gastélum-Garibaldi Jorge E. Sánchez-Rodríguez Jorge Arreola Electro-steric opening of the clc-2 chloride channel gate
description	Abstract The widely expressed two-pore homodimeric inward rectifier CLC-2 chloride channel regulates transepithelial chloride transport, extracellular chloride homeostasis, and neuronal excitability. Each pore is independently gated at hyperpolarized voltages by a conserved pore glutamate. Presumably, exiting chloride ions push glutamate outwardly while external protonation stabilizes it. To understand the mechanism of mouse CLC-2 opening we used homology modelling-guided structure–function analysis. Structural modelling suggests that glutamate E213 interacts with tyrosine Y561 to close a pore. Accordingly, Y561A and E213D mutants are activated at less hyperpolarized voltages, re-opened at depolarized voltages, and fast and common gating components are reduced. The double mutant cycle analysis showed that E213 and Y561 are energetically coupled to alter CLC-2 gating. In agreement, the anomalous mole fraction behaviour of the voltage dependence, measured by the voltage to induce half-open probability, was strongly altered in these mutants. Finally, cytosolic acidification or high extracellular chloride concentration, conditions that have little or no effect on WT CLC-2, induced reopening of Y561 mutants at positive voltages presumably by the inward opening of E213. We concluded that the CLC-2 gate is formed by Y561-E213 and that outward permeant anions open the gate by electrostatic and steric interactions.
format	article
author	José J. De Jesús-Pérez G. Arlette Méndez-Maldonado Ana E. López-Romero David Esparza-Jasso Irma L. González-Hernández Víctor De la Rosa Roberto Gastélum-Garibaldi Jorge E. Sánchez-Rodríguez Jorge Arreola
author_facet	José J. De Jesús-Pérez G. Arlette Méndez-Maldonado Ana E. López-Romero David Esparza-Jasso Irma L. González-Hernández Víctor De la Rosa Roberto Gastélum-Garibaldi Jorge E. Sánchez-Rodríguez Jorge Arreola
author_sort	José J. De Jesús-Pérez
title	Electro-steric opening of the clc-2 chloride channel gate
title_short	Electro-steric opening of the clc-2 chloride channel gate
title_full	Electro-steric opening of the clc-2 chloride channel gate
title_fullStr	Electro-steric opening of the clc-2 chloride channel gate
title_full_unstemmed	Electro-steric opening of the clc-2 chloride channel gate
title_sort	electro-steric opening of the clc-2 chloride channel gate
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/d68ca9a572f94156b3003b15ab5f24b7
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Electro-steric opening of the clc-2 chloride channel gate

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