Nicotinic receptor M3 transmembrane domain: Position 8′ contributes to channel gating

Autores
Rayes, Diego Hernán; de Rosa, Maria Jose; Spitzmaul, Guillermo Federico; Bouzat, Cecilia Beatriz
Año de publicación
2002
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The nicotinic acetylcholine receptor (nAChR) is a pentamer of homologous subunits with composition α2βεδ in adult muscle. Each subunit contains four transmembrane domains (M1-M4). Position 8′ of the M3 domain is phenylalanine in all heteromeric α subunits, whereas it is a hydrophobic nonaromatic residue in non-α subunits. Given this peculiar conservation pattern, we studied its contribution to muscle nAChR activation by combining mutagenesis with single-channel kinetic analysis. Construction of nAChRs carrying different numbers of phenylalanine residues at 8′ reveals that the mean open time decreases as a function of the number of phenylalanine residues. Thus, all subunits contribute through this position independently and additively to the channel closing rate. The impairment of channel opening increases when the number of phenylalanine residues at 8′ increases from two (wild-type nAChR) to five. The gating equilibrium constant of the latter mutant nAChR is 13-fold lower than that of the wild-type nAChR. The replacement of αF8′, βL8′, δV8′ and εV8′ by a series of hydrophobic amino acids reveals that the structural bases of the observed kinetic effects are nonequivalent among subunits. In the α subunit, hydrophobic amino acids at 8′ lead to prolonged channel lifetimes, whereas they lead either to normal kinetics (δ and ε subunits) or impaired channel gating (β subunit) in the non-α subunits. The overall results indicate that 8′ positions of the M3 domains of all subunits contribute to channel gating.
Fil: Rayes, Diego Hernán. 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: 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: 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
Materia
Nachr
Nicotinic Acetylcholine Receptor
Achr
Acetylcholine
M3
Third Transmembrane Domain
Hek
Human Embryonic Kidney
P Open
Channel Open Probability
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:11336/52910
Acceder
id CONICETDig_2553de52ab2cba4f441311cb65fbcb2a
oai_identifier_str oai:ri.conicet.gov.ar:11336/52910
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Nicotinic receptor M3 transmembrane domain: Position 8′ contributes to channel gatingRayes, Diego Hernánde Rosa, Maria JoseSpitzmaul, Guillermo FedericoBouzat, Cecilia BeatrizNachrNicotinic Acetylcholine ReceptorAchrAcetylcholineM3Third Transmembrane DomainHekHuman Embryonic KidneyP OpenChannel Open Probabilityhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The nicotinic acetylcholine receptor (nAChR) is a pentamer of homologous subunits with composition α2βεδ in adult muscle. Each subunit contains four transmembrane domains (M1-M4). Position 8′ of the M3 domain is phenylalanine in all heteromeric α subunits, whereas it is a hydrophobic nonaromatic residue in non-α subunits. Given this peculiar conservation pattern, we studied its contribution to muscle nAChR activation by combining mutagenesis with single-channel kinetic analysis. Construction of nAChRs carrying different numbers of phenylalanine residues at 8′ reveals that the mean open time decreases as a function of the number of phenylalanine residues. Thus, all subunits contribute through this position independently and additively to the channel closing rate. The impairment of channel opening increases when the number of phenylalanine residues at 8′ increases from two (wild-type nAChR) to five. The gating equilibrium constant of the latter mutant nAChR is 13-fold lower than that of the wild-type nAChR. The replacement of αF8′, βL8′, δV8′ and εV8′ by a series of hydrophobic amino acids reveals that the structural bases of the observed kinetic effects are nonequivalent among subunits. In the α subunit, hydrophobic amino acids at 8′ lead to prolonged channel lifetimes, whereas they lead either to normal kinetics (δ and ε subunits) or impaired channel gating (β subunit) in the non-α subunits. The overall results indicate that 8′ positions of the M3 domains of all subunits contribute to channel gating.Fil: Rayes, Diego Hernán. 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: 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: 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; ArgentinaAmerican Society for Pharmacology and Experimental Therapeutics2002-08-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/52910Rayes, Diego Hernán; de Rosa, Maria Jose; Spitzmaul, Guillermo Federico; Bouzat, Cecilia Beatriz; Nicotinic receptor M3 transmembrane domain: Position 8′ contributes to channel gating; American Society for Pharmacology and Experimental Therapeutics; Molecular Pharmacology; 62; 2; 1-8-2002; 406-4140026-895XCONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1124/mol.62.2.406info:eu-repo/semantics/altIdentifier/url/http://molpharm.aspetjournals.org/content/62/2/406info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:11:51Zoai:ri.conicet.gov.ar:11336/52910instacron: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:34982025-09-03 10:11:52.224CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Nicotinic receptor M3 transmembrane domain: Position 8′ contributes to channel gating
title Nicotinic receptor M3 transmembrane domain: Position 8′ contributes to channel gating
spellingShingle Nicotinic receptor M3 transmembrane domain: Position 8′ contributes to channel gating
Rayes, Diego Hernán
Nachr
Nicotinic Acetylcholine Receptor
Achr
Acetylcholine
M3
Third Transmembrane Domain
Hek
Human Embryonic Kidney
P Open
Channel Open Probability
title_short Nicotinic receptor M3 transmembrane domain: Position 8′ contributes to channel gating
title_full Nicotinic receptor M3 transmembrane domain: Position 8′ contributes to channel gating
title_fullStr Nicotinic receptor M3 transmembrane domain: Position 8′ contributes to channel gating
title_full_unstemmed Nicotinic receptor M3 transmembrane domain: Position 8′ contributes to channel gating
title_sort Nicotinic receptor M3 transmembrane domain: Position 8′ contributes to channel gating
dc.creator.none.fl_str_mv Rayes, Diego Hernán
de Rosa, Maria Jose
Spitzmaul, Guillermo Federico
Bouzat, Cecilia Beatriz
author Rayes, Diego Hernán
author_facet Rayes, Diego Hernán
de Rosa, Maria Jose
Spitzmaul, Guillermo Federico
Bouzat, Cecilia Beatriz
author_role author
author2 de Rosa, Maria Jose
Spitzmaul, Guillermo Federico
Bouzat, Cecilia Beatriz
author2_role author
author
author
dc.subject.none.fl_str_mv Nachr
Nicotinic Acetylcholine Receptor
Achr
Acetylcholine
M3
Third Transmembrane Domain
Hek
Human Embryonic Kidney
P Open
Channel Open Probability
topic Nachr
Nicotinic Acetylcholine Receptor
Achr
Acetylcholine
M3
Third Transmembrane Domain
Hek
Human Embryonic Kidney
P Open
Channel Open Probability
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 nicotinic acetylcholine receptor (nAChR) is a pentamer of homologous subunits with composition α2βεδ in adult muscle. Each subunit contains four transmembrane domains (M1-M4). Position 8′ of the M3 domain is phenylalanine in all heteromeric α subunits, whereas it is a hydrophobic nonaromatic residue in non-α subunits. Given this peculiar conservation pattern, we studied its contribution to muscle nAChR activation by combining mutagenesis with single-channel kinetic analysis. Construction of nAChRs carrying different numbers of phenylalanine residues at 8′ reveals that the mean open time decreases as a function of the number of phenylalanine residues. Thus, all subunits contribute through this position independently and additively to the channel closing rate. The impairment of channel opening increases when the number of phenylalanine residues at 8′ increases from two (wild-type nAChR) to five. The gating equilibrium constant of the latter mutant nAChR is 13-fold lower than that of the wild-type nAChR. The replacement of αF8′, βL8′, δV8′ and εV8′ by a series of hydrophobic amino acids reveals that the structural bases of the observed kinetic effects are nonequivalent among subunits. In the α subunit, hydrophobic amino acids at 8′ lead to prolonged channel lifetimes, whereas they lead either to normal kinetics (δ and ε subunits) or impaired channel gating (β subunit) in the non-α subunits. The overall results indicate that 8′ positions of the M3 domains of all subunits contribute to channel gating.
Fil: Rayes, Diego Hernán. 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: 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: 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 nicotinic acetylcholine receptor (nAChR) is a pentamer of homologous subunits with composition α2βεδ in adult muscle. Each subunit contains four transmembrane domains (M1-M4). Position 8′ of the M3 domain is phenylalanine in all heteromeric α subunits, whereas it is a hydrophobic nonaromatic residue in non-α subunits. Given this peculiar conservation pattern, we studied its contribution to muscle nAChR activation by combining mutagenesis with single-channel kinetic analysis. Construction of nAChRs carrying different numbers of phenylalanine residues at 8′ reveals that the mean open time decreases as a function of the number of phenylalanine residues. Thus, all subunits contribute through this position independently and additively to the channel closing rate. The impairment of channel opening increases when the number of phenylalanine residues at 8′ increases from two (wild-type nAChR) to five. The gating equilibrium constant of the latter mutant nAChR is 13-fold lower than that of the wild-type nAChR. The replacement of αF8′, βL8′, δV8′ and εV8′ by a series of hydrophobic amino acids reveals that the structural bases of the observed kinetic effects are nonequivalent among subunits. In the α subunit, hydrophobic amino acids at 8′ lead to prolonged channel lifetimes, whereas they lead either to normal kinetics (δ and ε subunits) or impaired channel gating (β subunit) in the non-α subunits. The overall results indicate that 8′ positions of the M3 domains of all subunits contribute to channel gating.
publishDate 2002
dc.date.none.fl_str_mv 2002-08-01
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/11336/52910
Rayes, Diego Hernán; de Rosa, Maria Jose; Spitzmaul, Guillermo Federico; Bouzat, Cecilia Beatriz; Nicotinic receptor M3 transmembrane domain: Position 8′ contributes to channel gating; American Society for Pharmacology and Experimental Therapeutics; Molecular Pharmacology; 62; 2; 1-8-2002; 406-414
0026-895X
CONICET Digital
CONICET
url http://hdl.handle.net/11336/52910
identifier_str_mv Rayes, Diego Hernán; de Rosa, Maria Jose; Spitzmaul, Guillermo Federico; Bouzat, Cecilia Beatriz; Nicotinic receptor M3 transmembrane domain: Position 8′ contributes to channel gating; American Society for Pharmacology and Experimental Therapeutics; Molecular Pharmacology; 62; 2; 1-8-2002; 406-414
0026-895X
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1124/mol.62.2.406
info:eu-repo/semantics/altIdentifier/url/http://molpharm.aspetjournals.org/content/62/2/406
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
dc.publisher.none.fl_str_mv American Society for Pharmacology and Experimental Therapeutics
publisher.none.fl_str_mv American Society for Pharmacology and Experimental Therapeutics
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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score 13.13397

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