Role of Pairwise Interactions between M1 and M2 Domains of the Nicotinic Receptor in Channel Gating

Autores
Corradi, Jeremias; Spitzmaul, Guillermo Federico; de Rosa, Maria Jose; Costabel, Marcelo Daniel; Bouzat, Cecilia Beatriz
Año de publicación
2007
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The adult form of the nicotinic acetylcholine receptor (AChR) consists of five subunits (α2βɛδ), each having four transmembrane domains (M1–M4). The atomic model of the nicotinic acetylcholine receptor shows that the pore-lining M2 domains make no extensive contacts with the rest of the transmembrane domains. However, there are several sites where close appositions between segments occur. It has been suggested that the pair αM1-F15′ and αM2-L11′ is one of the potential interactions between segments. To determine experimentally if these residues are interacting and to explore if this interhelical interaction is essential for channel gating, we combined mutagenesis with single-channel kinetic analysis. Mutations in αM1-F15′ lead to profound changes in the opening rate and slighter changes in the closing rate. Channel gating is impaired as the volume of the residue increases. Rate-equilibrium linear free-energy relationship analysis reveals an ∼70% open-state-like environment for αM1-F15′ at the transition state of the gating reaction, suggesting that it moves early during the gating process. Replacing the residue at αM1-15′ by that at αM2-11′ and vice versa profoundly alters gating, but the combination of the two mutations restores gating to near normal, indicating that αM1-F15′ and αM2-L11′ are interchangeable. Double-mutant cycle analysis shows that these residues are energetically coupled. Thus, the interaction between M1 and M2 plays a key role in channel gating.
Fil: Corradi, Jeremias. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: Spitzmaul, Guillermo Federico. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: de Rosa, Maria Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: Costabel, Marcelo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional del Sur. Departamento de Física; Argentina
Fil: Bouzat, Cecilia Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/44405
Acceder
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network_name_str CONICET Digital (CONICET)
spelling Role of Pairwise Interactions between M1 and M2 Domains of the Nicotinic Receptor in Channel GatingCorradi, JeremiasSpitzmaul, Guillermo Federicode Rosa, Maria JoseCostabel, Marcelo DanielBouzat, Cecilia Beatrizhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The adult form of the nicotinic acetylcholine receptor (AChR) consists of five subunits (α2βɛδ), each having four transmembrane domains (M1–M4). The atomic model of the nicotinic acetylcholine receptor shows that the pore-lining M2 domains make no extensive contacts with the rest of the transmembrane domains. However, there are several sites where close appositions between segments occur. It has been suggested that the pair αM1-F15′ and αM2-L11′ is one of the potential interactions between segments. To determine experimentally if these residues are interacting and to explore if this interhelical interaction is essential for channel gating, we combined mutagenesis with single-channel kinetic analysis. Mutations in αM1-F15′ lead to profound changes in the opening rate and slighter changes in the closing rate. Channel gating is impaired as the volume of the residue increases. Rate-equilibrium linear free-energy relationship analysis reveals an ∼70% open-state-like environment for αM1-F15′ at the transition state of the gating reaction, suggesting that it moves early during the gating process. Replacing the residue at αM1-15′ by that at αM2-11′ and vice versa profoundly alters gating, but the combination of the two mutations restores gating to near normal, indicating that αM1-F15′ and αM2-L11′ are interchangeable. Double-mutant cycle analysis shows that these residues are energetically coupled. Thus, the interaction between M1 and M2 plays a key role in channel gating.Fil: Corradi, Jeremias. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; ArgentinaFil: Spitzmaul, Guillermo Federico. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; ArgentinaFil: de Rosa, Maria Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; ArgentinaFil: Costabel, Marcelo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional del Sur. Departamento de Física; ArgentinaFil: Bouzat, Cecilia Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; ArgentinaCell Press2007-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/44405Corradi, Jeremias; Spitzmaul, Guillermo Federico; de Rosa, Maria Jose; Costabel, Marcelo Daniel; Bouzat, Cecilia Beatriz; Role of Pairwise Interactions between M1 and M2 Domains of the Nicotinic Receptor in Channel Gating; Cell Press; Biophysical Journal; 92; 1; 1-2007; 76-860006-3495CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1697868/info:eu-repo/semantics/altIdentifier/doi/10.1529/biophysj.106.088757info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0006349507708068info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:45:15Zoai:ri.conicet.gov.ar:11336/44405instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 09:45:15.717CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Role of Pairwise Interactions between M1 and M2 Domains of the Nicotinic Receptor in Channel Gating
title Role of Pairwise Interactions between M1 and M2 Domains of the Nicotinic Receptor in Channel Gating
spellingShingle Role of Pairwise Interactions between M1 and M2 Domains of the Nicotinic Receptor in Channel Gating
Corradi, Jeremias
title_short Role of Pairwise Interactions between M1 and M2 Domains of the Nicotinic Receptor in Channel Gating
title_full Role of Pairwise Interactions between M1 and M2 Domains of the Nicotinic Receptor in Channel Gating
title_fullStr Role of Pairwise Interactions between M1 and M2 Domains of the Nicotinic Receptor in Channel Gating
title_full_unstemmed Role of Pairwise Interactions between M1 and M2 Domains of the Nicotinic Receptor in Channel Gating
title_sort Role of Pairwise Interactions between M1 and M2 Domains of the Nicotinic Receptor in Channel Gating
dc.creator.none.fl_str_mv Corradi, Jeremias
Spitzmaul, Guillermo Federico
de Rosa, Maria Jose
Costabel, Marcelo Daniel
Bouzat, Cecilia Beatriz
author Corradi, Jeremias
author_facet Corradi, Jeremias
Spitzmaul, Guillermo Federico
de Rosa, Maria Jose
Costabel, Marcelo Daniel
Bouzat, Cecilia Beatriz
author_role author
author2 Spitzmaul, Guillermo Federico
de Rosa, Maria Jose
Costabel, Marcelo Daniel
Bouzat, Cecilia Beatriz
author2_role author
author
author
author
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv The adult form of the nicotinic acetylcholine receptor (AChR) consists of five subunits (α2βɛδ), each having four transmembrane domains (M1–M4). The atomic model of the nicotinic acetylcholine receptor shows that the pore-lining M2 domains make no extensive contacts with the rest of the transmembrane domains. However, there are several sites where close appositions between segments occur. It has been suggested that the pair αM1-F15′ and αM2-L11′ is one of the potential interactions between segments. To determine experimentally if these residues are interacting and to explore if this interhelical interaction is essential for channel gating, we combined mutagenesis with single-channel kinetic analysis. Mutations in αM1-F15′ lead to profound changes in the opening rate and slighter changes in the closing rate. Channel gating is impaired as the volume of the residue increases. Rate-equilibrium linear free-energy relationship analysis reveals an ∼70% open-state-like environment for αM1-F15′ at the transition state of the gating reaction, suggesting that it moves early during the gating process. Replacing the residue at αM1-15′ by that at αM2-11′ and vice versa profoundly alters gating, but the combination of the two mutations restores gating to near normal, indicating that αM1-F15′ and αM2-L11′ are interchangeable. Double-mutant cycle analysis shows that these residues are energetically coupled. Thus, the interaction between M1 and M2 plays a key role in channel gating.
Fil: Corradi, Jeremias. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: Spitzmaul, Guillermo Federico. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: de Rosa, Maria Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: Costabel, Marcelo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional del Sur. Departamento de Física; Argentina
Fil: Bouzat, Cecilia Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
description The adult form of the nicotinic acetylcholine receptor (AChR) consists of five subunits (α2βɛδ), each having four transmembrane domains (M1–M4). The atomic model of the nicotinic acetylcholine receptor shows that the pore-lining M2 domains make no extensive contacts with the rest of the transmembrane domains. However, there are several sites where close appositions between segments occur. It has been suggested that the pair αM1-F15′ and αM2-L11′ is one of the potential interactions between segments. To determine experimentally if these residues are interacting and to explore if this interhelical interaction is essential for channel gating, we combined mutagenesis with single-channel kinetic analysis. Mutations in αM1-F15′ lead to profound changes in the opening rate and slighter changes in the closing rate. Channel gating is impaired as the volume of the residue increases. Rate-equilibrium linear free-energy relationship analysis reveals an ∼70% open-state-like environment for αM1-F15′ at the transition state of the gating reaction, suggesting that it moves early during the gating process. Replacing the residue at αM1-15′ by that at αM2-11′ and vice versa profoundly alters gating, but the combination of the two mutations restores gating to near normal, indicating that αM1-F15′ and αM2-L11′ are interchangeable. Double-mutant cycle analysis shows that these residues are energetically coupled. Thus, the interaction between M1 and M2 plays a key role in channel gating.
publishDate 2007
dc.date.none.fl_str_mv 2007-01
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/44405
Corradi, Jeremias; Spitzmaul, Guillermo Federico; de Rosa, Maria Jose; Costabel, Marcelo Daniel; Bouzat, Cecilia Beatriz; Role of Pairwise Interactions between M1 and M2 Domains of the Nicotinic Receptor in Channel Gating; Cell Press; Biophysical Journal; 92; 1; 1-2007; 76-86
0006-3495
CONICET Digital
CONICET
url http://hdl.handle.net/11336/44405
identifier_str_mv Corradi, Jeremias; Spitzmaul, Guillermo Federico; de Rosa, Maria Jose; Costabel, Marcelo Daniel; Bouzat, Cecilia Beatriz; Role of Pairwise Interactions between M1 and M2 Domains of the Nicotinic Receptor in Channel Gating; Cell Press; Biophysical Journal; 92; 1; 1-2007; 76-86
0006-3495
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1697868/
info:eu-repo/semantics/altIdentifier/doi/10.1529/biophysj.106.088757
info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0006349507708068
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Cell Press
publisher.none.fl_str_mv Cell Press
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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