Phosphorylation of intracellular tyrosines modulates the ionotropic function of the α7 nicotinic receptor

Autores
Chrestia, Juan Facundo; Bouzat, Cecilia Beatriz; Esandi, María del Carmen
Año de publicación
2018
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
α7 is expressed in the brain and contributes to cognition, attention, and memory. It contains an extracellular domain with the agonist binding sites; a transmembrane domain, which forms the ion pore; and an intracellular domain (ICD), which contains sites for modulation and intracellular signaling. The mechanisms by which the cell can regulate the ionotropic function of α7 remain unknown. We explored how intracellular phosphorylation affects α7 activity by patch clamp recordings in HEK cells expressing α7. Wild-type α7 channel activity elicited by ACh appears as brief isolated openings and as activation episodes containing a few brief openings in quick succession (bursts). Preincubation of cells expressing α7 with the inhibitor of Src family kinases (PP2) increased significantly the mean burst duration. The exposure of cells to PP2 during the course of the recording revealed a significant increase in the frequency of channel opening in addition to the increase of burst durations. To confirm that these changes were due to the inhibition of phosphorylation of α7-ICD, we introduced mutations at potential phosphorylation sites (Y386F and Y442F). The mutations prolonged burst durations, thus mimicking the effects of PP2. Also, the mutants were insensitive to PP2, confirming that Y386 and Y442 are responsible for its effects on α7 kinetics. Our results indicate that dephosphorylation positively modulates α7 channel activity in a way compatible with decreased desensitization.
Fil: Chrestia, Juan Facundo. 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. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; 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. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
Fil: Esandi, María del Carmen. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina. 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
XXXIII Congreso Anual de la Sociedad Argentina de Investigación en Neurociencias
Córdoba
Argentina
Sociedad Argentina de Investigación en Neurociencias
Materia
NICOTINIC RECEPTOR
TYROSYNES
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:11336/232189
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/232189
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Phosphorylation of intracellular tyrosines modulates the ionotropic function of the α7 nicotinic receptorChrestia, Juan FacundoBouzat, Cecilia BeatrizEsandi, María del CarmenNICOTINIC RECEPTORTYROSYNEShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1α7 is expressed in the brain and contributes to cognition, attention, and memory. It contains an extracellular domain with the agonist binding sites; a transmembrane domain, which forms the ion pore; and an intracellular domain (ICD), which contains sites for modulation and intracellular signaling. The mechanisms by which the cell can regulate the ionotropic function of α7 remain unknown. We explored how intracellular phosphorylation affects α7 activity by patch clamp recordings in HEK cells expressing α7. Wild-type α7 channel activity elicited by ACh appears as brief isolated openings and as activation episodes containing a few brief openings in quick succession (bursts). Preincubation of cells expressing α7 with the inhibitor of Src family kinases (PP2) increased significantly the mean burst duration. The exposure of cells to PP2 during the course of the recording revealed a significant increase in the frequency of channel opening in addition to the increase of burst durations. To confirm that these changes were due to the inhibition of phosphorylation of α7-ICD, we introduced mutations at potential phosphorylation sites (Y386F and Y442F). The mutations prolonged burst durations, thus mimicking the effects of PP2. Also, the mutants were insensitive to PP2, confirming that Y386 and Y442 are responsible for its effects on α7 kinetics. Our results indicate that dephosphorylation positively modulates α7 channel activity in a way compatible with decreased desensitization.Fil: Chrestia, Juan Facundo. 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. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; 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; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; ArgentinaFil: Esandi, María del Carmen. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina. 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; ArgentinaXXXIII Congreso Anual de la Sociedad Argentina de Investigación en NeurocienciasCórdobaArgentinaSociedad Argentina de Investigación en NeurocienciasSociedad Argentina de Investigación en Neurociencias2018info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectCongresoBookhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/vnd.openxmlformats-officedocument.wordprocessingml.documentapplication/pdfhttp://hdl.handle.net/11336/232189Phosphorylation of intracellular tyrosines modulates the ionotropic function of the α7 nicotinic receptor; XXXIII Congreso Anual de la Sociedad Argentina de Investigación en Neurociencias; Córdoba; Argentina; 2018; 312-312CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://saneurociencias.org.ar/congresos-san-2/Nacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:38:57Zoai:ri.conicet.gov.ar:11336/232189instacron: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 10:38:57.288CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Phosphorylation of intracellular tyrosines modulates the ionotropic function of the α7 nicotinic receptor
title Phosphorylation of intracellular tyrosines modulates the ionotropic function of the α7 nicotinic receptor
spellingShingle Phosphorylation of intracellular tyrosines modulates the ionotropic function of the α7 nicotinic receptor
Chrestia, Juan Facundo
NICOTINIC RECEPTOR
TYROSYNES
title_short Phosphorylation of intracellular tyrosines modulates the ionotropic function of the α7 nicotinic receptor
title_full Phosphorylation of intracellular tyrosines modulates the ionotropic function of the α7 nicotinic receptor
title_fullStr Phosphorylation of intracellular tyrosines modulates the ionotropic function of the α7 nicotinic receptor
title_full_unstemmed Phosphorylation of intracellular tyrosines modulates the ionotropic function of the α7 nicotinic receptor
title_sort Phosphorylation of intracellular tyrosines modulates the ionotropic function of the α7 nicotinic receptor
dc.creator.none.fl_str_mv Chrestia, Juan Facundo
Bouzat, Cecilia Beatriz
Esandi, María del Carmen
author Chrestia, Juan Facundo
author_facet Chrestia, Juan Facundo
Bouzat, Cecilia Beatriz
Esandi, María del Carmen
author_role author
author2 Bouzat, Cecilia Beatriz
Esandi, María del Carmen
author2_role author
author
dc.subject.none.fl_str_mv NICOTINIC RECEPTOR
TYROSYNES
topic NICOTINIC RECEPTOR
TYROSYNES
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv α7 is expressed in the brain and contributes to cognition, attention, and memory. It contains an extracellular domain with the agonist binding sites; a transmembrane domain, which forms the ion pore; and an intracellular domain (ICD), which contains sites for modulation and intracellular signaling. The mechanisms by which the cell can regulate the ionotropic function of α7 remain unknown. We explored how intracellular phosphorylation affects α7 activity by patch clamp recordings in HEK cells expressing α7. Wild-type α7 channel activity elicited by ACh appears as brief isolated openings and as activation episodes containing a few brief openings in quick succession (bursts). Preincubation of cells expressing α7 with the inhibitor of Src family kinases (PP2) increased significantly the mean burst duration. The exposure of cells to PP2 during the course of the recording revealed a significant increase in the frequency of channel opening in addition to the increase of burst durations. To confirm that these changes were due to the inhibition of phosphorylation of α7-ICD, we introduced mutations at potential phosphorylation sites (Y386F and Y442F). The mutations prolonged burst durations, thus mimicking the effects of PP2. Also, the mutants were insensitive to PP2, confirming that Y386 and Y442 are responsible for its effects on α7 kinetics. Our results indicate that dephosphorylation positively modulates α7 channel activity in a way compatible with decreased desensitization.
Fil: Chrestia, Juan Facundo. 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. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; 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. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
Fil: Esandi, María del Carmen. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina. 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
XXXIII Congreso Anual de la Sociedad Argentina de Investigación en Neurociencias
Córdoba
Argentina
Sociedad Argentina de Investigación en Neurociencias
description α7 is expressed in the brain and contributes to cognition, attention, and memory. It contains an extracellular domain with the agonist binding sites; a transmembrane domain, which forms the ion pore; and an intracellular domain (ICD), which contains sites for modulation and intracellular signaling. The mechanisms by which the cell can regulate the ionotropic function of α7 remain unknown. We explored how intracellular phosphorylation affects α7 activity by patch clamp recordings in HEK cells expressing α7. Wild-type α7 channel activity elicited by ACh appears as brief isolated openings and as activation episodes containing a few brief openings in quick succession (bursts). Preincubation of cells expressing α7 with the inhibitor of Src family kinases (PP2) increased significantly the mean burst duration. The exposure of cells to PP2 during the course of the recording revealed a significant increase in the frequency of channel opening in addition to the increase of burst durations. To confirm that these changes were due to the inhibition of phosphorylation of α7-ICD, we introduced mutations at potential phosphorylation sites (Y386F and Y442F). The mutations prolonged burst durations, thus mimicking the effects of PP2. Also, the mutants were insensitive to PP2, confirming that Y386 and Y442 are responsible for its effects on α7 kinetics. Our results indicate that dephosphorylation positively modulates α7 channel activity in a way compatible with decreased desensitization.
publishDate 2018
dc.date.none.fl_str_mv 2018
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Congreso
Book
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/232189
Phosphorylation of intracellular tyrosines modulates the ionotropic function of the α7 nicotinic receptor; XXXIII Congreso Anual de la Sociedad Argentina de Investigación en Neurociencias; Córdoba; Argentina; 2018; 312-312
CONICET Digital
CONICET
url http://hdl.handle.net/11336/232189
identifier_str_mv Phosphorylation of intracellular tyrosines modulates the ionotropic function of the α7 nicotinic receptor; XXXIII Congreso Anual de la Sociedad Argentina de Investigación en Neurociencias; Córdoba; Argentina; 2018; 312-312
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://saneurociencias.org.ar/congresos-san-2/
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/
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application/vnd.openxmlformats-officedocument.wordprocessingml.document
application/pdf
dc.coverage.none.fl_str_mv Nacional
dc.publisher.none.fl_str_mv Sociedad Argentina de Investigación en Neurociencias
publisher.none.fl_str_mv Sociedad Argentina de Investigación en Neurociencias
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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