Equilibrium unfolding of the PDZ domain of β2-syntrophin
- Autores
- Torchio, Gabriela María; Ermacora, Mario Roberto; Sica, Mauricio Pablo
- Año de publicación
- 2012
- Idioma
- inglés
- Tipo de recurso
- artículo
- Estado
- versión publicada
- Descripción
- b2-syntrophin, a dystrophin-associated protein, plays a pivotal role in insulin secretion by pancreatic b-cells. Itcontains a PDZ domain (b2S-PDZ) that, in complex with protein-tyrosine phosphatase ICA512, anchors the dense insulin gran-ules to actin filaments. The phosphorylation state of b2-syntrophin allosterically regulates the affinity of b2S-PDZ for ICA512,and the disruption of the complex triggers the mobilization of the insulin granule stores. Here, we investigate the thermal unfold-ing of b2S-PDZ at different pH and urea concentrations. Our results indicate that, unlike other PDZ domains, b2S-PDZ is margin-ally stable. Thermal denaturation experiments show broad transitions and cold denaturation, and a two-state model fit revealsa significant unfolded fraction under physiological conditions. Furthermore, Tm and Tmax denaturant-dependent shifts andnoncoincidence of melting curves monitored at different wavelengths suggest that two-state and three-state models fail toexplain the equilibrium data properly and are in better agreement with a downhill scenario. Its higher stability at pH >9 andthe results of molecular dynamics simulations indicate that this behavior of b2S-PDZ might be related to its charge distribution.All together, our results suggest a link between the conformational plasticity of the native ensemble of this PDZ domain and theregulation of insulin secretion.
Fil: Torchio, Gabriela María. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Expresion y Plegamiento de Proteinas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Ermacora, Mario Roberto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Expresion y Plegamiento de Proteinas; Argentina
Fil: Sica, Mauricio Pablo. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Expresion y Plegamiento de Proteinas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina - Materia
-
syntrophin
PDZ
unfolding
downhill - 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:11336/196602
Ver los metadatos del registro completo
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Equilibrium unfolding of the PDZ domain of β2-syntrophinTorchio, Gabriela MaríaErmacora, Mario RobertoSica, Mauricio PablosyntrophinPDZunfoldingdownhillhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1b2-syntrophin, a dystrophin-associated protein, plays a pivotal role in insulin secretion by pancreatic b-cells. Itcontains a PDZ domain (b2S-PDZ) that, in complex with protein-tyrosine phosphatase ICA512, anchors the dense insulin gran-ules to actin filaments. The phosphorylation state of b2-syntrophin allosterically regulates the affinity of b2S-PDZ for ICA512,and the disruption of the complex triggers the mobilization of the insulin granule stores. Here, we investigate the thermal unfold-ing of b2S-PDZ at different pH and urea concentrations. Our results indicate that, unlike other PDZ domains, b2S-PDZ is margin-ally stable. Thermal denaturation experiments show broad transitions and cold denaturation, and a two-state model fit revealsa significant unfolded fraction under physiological conditions. Furthermore, Tm and Tmax denaturant-dependent shifts andnoncoincidence of melting curves monitored at different wavelengths suggest that two-state and three-state models fail toexplain the equilibrium data properly and are in better agreement with a downhill scenario. Its higher stability at pH >9 andthe results of molecular dynamics simulations indicate that this behavior of b2S-PDZ might be related to its charge distribution.All together, our results suggest a link between the conformational plasticity of the native ensemble of this PDZ domain and theregulation of insulin secretion.Fil: Torchio, Gabriela María. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Expresion y Plegamiento de Proteinas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Ermacora, Mario Roberto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Expresion y Plegamiento de Proteinas; ArgentinaFil: Sica, Mauricio Pablo. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Expresion y Plegamiento de Proteinas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaCell Press2012-06info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/196602Torchio, Gabriela María; Ermacora, Mario Roberto; Sica, Mauricio Pablo; Equilibrium unfolding of the PDZ domain of β2-syntrophin; Cell Press; Biophysical Journal; 102; 12; 6-2012; 2835-28440006-3495CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0006349512005747info:eu-repo/semantics/altIdentifier/doi/10.1016/j.bpj.2012.05.021info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:11:19Zoai:ri.conicet.gov.ar:11336/196602instacron: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 10:11:19.755CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse |
dc.title.none.fl_str_mv |
Equilibrium unfolding of the PDZ domain of β2-syntrophin |
title |
Equilibrium unfolding of the PDZ domain of β2-syntrophin |
spellingShingle |
Equilibrium unfolding of the PDZ domain of β2-syntrophin Torchio, Gabriela María syntrophin PDZ unfolding downhill |
title_short |
Equilibrium unfolding of the PDZ domain of β2-syntrophin |
title_full |
Equilibrium unfolding of the PDZ domain of β2-syntrophin |
title_fullStr |
Equilibrium unfolding of the PDZ domain of β2-syntrophin |
title_full_unstemmed |
Equilibrium unfolding of the PDZ domain of β2-syntrophin |
title_sort |
Equilibrium unfolding of the PDZ domain of β2-syntrophin |
dc.creator.none.fl_str_mv |
Torchio, Gabriela María Ermacora, Mario Roberto Sica, Mauricio Pablo |
author |
Torchio, Gabriela María |
author_facet |
Torchio, Gabriela María Ermacora, Mario Roberto Sica, Mauricio Pablo |
author_role |
author |
author2 |
Ermacora, Mario Roberto Sica, Mauricio Pablo |
author2_role |
author author |
dc.subject.none.fl_str_mv |
syntrophin PDZ unfolding downhill |
topic |
syntrophin PDZ unfolding downhill |
purl_subject.fl_str_mv |
https://purl.org/becyt/ford/1.6 https://purl.org/becyt/ford/1 |
dc.description.none.fl_txt_mv |
b2-syntrophin, a dystrophin-associated protein, plays a pivotal role in insulin secretion by pancreatic b-cells. Itcontains a PDZ domain (b2S-PDZ) that, in complex with protein-tyrosine phosphatase ICA512, anchors the dense insulin gran-ules to actin filaments. The phosphorylation state of b2-syntrophin allosterically regulates the affinity of b2S-PDZ for ICA512,and the disruption of the complex triggers the mobilization of the insulin granule stores. Here, we investigate the thermal unfold-ing of b2S-PDZ at different pH and urea concentrations. Our results indicate that, unlike other PDZ domains, b2S-PDZ is margin-ally stable. Thermal denaturation experiments show broad transitions and cold denaturation, and a two-state model fit revealsa significant unfolded fraction under physiological conditions. Furthermore, Tm and Tmax denaturant-dependent shifts andnoncoincidence of melting curves monitored at different wavelengths suggest that two-state and three-state models fail toexplain the equilibrium data properly and are in better agreement with a downhill scenario. Its higher stability at pH >9 andthe results of molecular dynamics simulations indicate that this behavior of b2S-PDZ might be related to its charge distribution.All together, our results suggest a link between the conformational plasticity of the native ensemble of this PDZ domain and theregulation of insulin secretion. Fil: Torchio, Gabriela María. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Expresion y Plegamiento de Proteinas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Ermacora, Mario Roberto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Expresion y Plegamiento de Proteinas; Argentina Fil: Sica, Mauricio Pablo. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Expresion y Plegamiento de Proteinas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina |
description |
b2-syntrophin, a dystrophin-associated protein, plays a pivotal role in insulin secretion by pancreatic b-cells. Itcontains a PDZ domain (b2S-PDZ) that, in complex with protein-tyrosine phosphatase ICA512, anchors the dense insulin gran-ules to actin filaments. The phosphorylation state of b2-syntrophin allosterically regulates the affinity of b2S-PDZ for ICA512,and the disruption of the complex triggers the mobilization of the insulin granule stores. Here, we investigate the thermal unfold-ing of b2S-PDZ at different pH and urea concentrations. Our results indicate that, unlike other PDZ domains, b2S-PDZ is margin-ally stable. Thermal denaturation experiments show broad transitions and cold denaturation, and a two-state model fit revealsa significant unfolded fraction under physiological conditions. Furthermore, Tm and Tmax denaturant-dependent shifts andnoncoincidence of melting curves monitored at different wavelengths suggest that two-state and three-state models fail toexplain the equilibrium data properly and are in better agreement with a downhill scenario. Its higher stability at pH >9 andthe results of molecular dynamics simulations indicate that this behavior of b2S-PDZ might be related to its charge distribution.All together, our results suggest a link between the conformational plasticity of the native ensemble of this PDZ domain and theregulation of insulin secretion. |
publishDate |
2012 |
dc.date.none.fl_str_mv |
2012-06 |
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/11336/196602 Torchio, Gabriela María; Ermacora, Mario Roberto; Sica, Mauricio Pablo; Equilibrium unfolding of the PDZ domain of β2-syntrophin; Cell Press; Biophysical Journal; 102; 12; 6-2012; 2835-2844 0006-3495 CONICET Digital CONICET |
url |
http://hdl.handle.net/11336/196602 |
identifier_str_mv |
Torchio, Gabriela María; Ermacora, Mario Roberto; Sica, Mauricio Pablo; Equilibrium unfolding of the PDZ domain of β2-syntrophin; Cell Press; Biophysical Journal; 102; 12; 6-2012; 2835-2844 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.sciencedirect.com/science/article/pii/S0006349512005747 info:eu-repo/semantics/altIdentifier/doi/10.1016/j.bpj.2012.05.021 |
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 |
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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1844614011243462656 |
score |
13.070432 |