Mutational analysis to explore long-range allosteric couplings involved in a pentameric channel receptor pre-activation and activation

Pentameric ligand-gated ion channels (pLGICs) mediate chemical signaling through a succession of allosteric transitions that are yet not completely understood as intermediate states remain poorly characterized by structural approaches. In a previous study on the prototypic bacterial proton-gated cha...

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Autores principales:	Solène N Lefebvre, Antoine Taly, Anaïs Menny, Karima Medjebeur, Pierre-Jean Corringer
Formato:	article
Lenguaje:	EN
Publicado:	eLife Sciences Publications Ltd 2021
Materias:	fluorescence quenching pentameric ligand-gated ion channels intermediate conformation allosteric coupling Medicine R Science Q Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/47b01ac4ec284c219fa466c9ed105727
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spelling	oai:doaj.org-article:47b01ac4ec284c219fa466c9ed1057272021-11-23T13:03:23ZMutational analysis to explore long-range allosteric couplings involved in a pentameric channel receptor pre-activation and activation10.7554/eLife.606822050-084Xe60682https://doaj.org/article/47b01ac4ec284c219fa466c9ed1057272021-09-01T00:00:00Zhttps://elifesciences.org/articles/60682https://doaj.org/toc/2050-084XPentameric ligand-gated ion channels (pLGICs) mediate chemical signaling through a succession of allosteric transitions that are yet not completely understood as intermediate states remain poorly characterized by structural approaches. In a previous study on the prototypic bacterial proton-gated channel GLIC, we generated several fluorescent sensors of the protein conformation that report a fast transition to a pre-active state, which precedes the slower process of activation with pore opening. Here, we explored the phenotype of a series of allosteric mutations, using simultaneous steady-state fluorescence and electrophysiological measurements over a broad pH range. Our data, fitted to a three-state Monod-Wyman-Changeux model, show that mutations at the subunit interface in the extracellular domain (ECD) principally alter pre-activation, while mutations in the lower ECD and in the transmembrane domain principally alter activation. We also show that propofol alters both transitions. Data are discussed in the framework of transition pathways generated by normal mode analysis (iModFit). It further supports that pre-activation involves major quaternary compaction of the ECD, and suggests that activation involves principally a reorganization of a ‘central gating region’ involving a contraction of the ECD β-sandwich and the tilt of the channel lining M2 helix.Solène N LefebvreAntoine TalyAnaïs MennyKarima MedjebeurPierre-Jean CorringereLife Sciences Publications Ltdarticlefluorescence quenchingpentameric ligand-gated ion channelsintermediate conformationallosteric couplingMedicineRScienceQBiology (General)QH301-705.5ENeLife, Vol 10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	fluorescence quenching pentameric ligand-gated ion channels intermediate conformation allosteric coupling Medicine R Science Q Biology (General) QH301-705.5
spellingShingle	fluorescence quenching pentameric ligand-gated ion channels intermediate conformation allosteric coupling Medicine R Science Q Biology (General) QH301-705.5 Solène N Lefebvre Antoine Taly Anaïs Menny Karima Medjebeur Pierre-Jean Corringer Mutational analysis to explore long-range allosteric couplings involved in a pentameric channel receptor pre-activation and activation
description	Pentameric ligand-gated ion channels (pLGICs) mediate chemical signaling through a succession of allosteric transitions that are yet not completely understood as intermediate states remain poorly characterized by structural approaches. In a previous study on the prototypic bacterial proton-gated channel GLIC, we generated several fluorescent sensors of the protein conformation that report a fast transition to a pre-active state, which precedes the slower process of activation with pore opening. Here, we explored the phenotype of a series of allosteric mutations, using simultaneous steady-state fluorescence and electrophysiological measurements over a broad pH range. Our data, fitted to a three-state Monod-Wyman-Changeux model, show that mutations at the subunit interface in the extracellular domain (ECD) principally alter pre-activation, while mutations in the lower ECD and in the transmembrane domain principally alter activation. We also show that propofol alters both transitions. Data are discussed in the framework of transition pathways generated by normal mode analysis (iModFit). It further supports that pre-activation involves major quaternary compaction of the ECD, and suggests that activation involves principally a reorganization of a ‘central gating region’ involving a contraction of the ECD β-sandwich and the tilt of the channel lining M2 helix.
format	article
author	Solène N Lefebvre Antoine Taly Anaïs Menny Karima Medjebeur Pierre-Jean Corringer
author_facet	Solène N Lefebvre Antoine Taly Anaïs Menny Karima Medjebeur Pierre-Jean Corringer
author_sort	Solène N Lefebvre
title	Mutational analysis to explore long-range allosteric couplings involved in a pentameric channel receptor pre-activation and activation
title_short	Mutational analysis to explore long-range allosteric couplings involved in a pentameric channel receptor pre-activation and activation
title_full	Mutational analysis to explore long-range allosteric couplings involved in a pentameric channel receptor pre-activation and activation
title_fullStr	Mutational analysis to explore long-range allosteric couplings involved in a pentameric channel receptor pre-activation and activation
title_full_unstemmed	Mutational analysis to explore long-range allosteric couplings involved in a pentameric channel receptor pre-activation and activation
title_sort	mutational analysis to explore long-range allosteric couplings involved in a pentameric channel receptor pre-activation and activation
publisher	eLife Sciences Publications Ltd
publishDate	2021
url	https://doaj.org/article/47b01ac4ec284c219fa466c9ed105727
work_keys_str_mv	AT solenenlefebvre mutationalanalysistoexplorelongrangeallostericcouplingsinvolvedinapentamericchannelreceptorpreactivationandactivation AT antoinetaly mutationalanalysistoexplorelongrangeallostericcouplingsinvolvedinapentamericchannelreceptorpreactivationandactivation AT anaismenny mutationalanalysistoexplorelongrangeallostericcouplingsinvolvedinapentamericchannelreceptorpreactivationandactivation AT karimamedjebeur mutationalanalysistoexplorelongrangeallostericcouplingsinvolvedinapentamericchannelreceptorpreactivationandactivation AT pierrejeancorringer mutationalanalysistoexplorelongrangeallostericcouplingsinvolvedinapentamericchannelreceptorpreactivationandactivation
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Mutational analysis to explore long-range allosteric couplings involved in a pentameric channel receptor pre-activation and activation

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