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
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/232189
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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/ |
dc.format.none.fl_str_mv |
application/pdf 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 |
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reponame:CONICET Digital (CONICET) instname:Consejo Nacional de Investigaciones Científicas y Técnicas |
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Consejo Nacional de Investigaciones Científicas y Técnicas |
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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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