Equilibrium unfolding of the PDZ domain of β2-syntrophin

β2-syntrophin, a dystrophin-associated protein, plays a pivotal role in insulin secretion by pancreatic β-cells. It contains a PDZ domain (β2S-PDZ) that, in complex with protein-tyrosine phosphatase ICA512, anchors the dense insulin granules to actin filaments. The phosphorylation state of β2-syntro...

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Autores principales: Torchio, Gabriela María, Ermácora, Mario Roberto, Sica, Mauricio Pablo
Formato: Articulo
Lenguaje:Inglés
Publicado: 2012
Materias:
Acceso en línea:http://sedici.unlp.edu.ar/handle/10915/87841
Aporte de:
id I19-R120-10915-87841
record_format dspace
institution Universidad Nacional de La Plata
institution_str I-19
repository_str R-120
collection SEDICI (UNLP)
language Inglés
topic Ciencias Exactas
β2-syntrophin
PDZ domain
spellingShingle Ciencias Exactas
β2-syntrophin
PDZ domain
Torchio, Gabriela María
Ermácora, Mario Roberto
Sica, Mauricio Pablo
Equilibrium unfolding of the PDZ domain of β2-syntrophin
topic_facet Ciencias Exactas
β2-syntrophin
PDZ domain
description β2-syntrophin, a dystrophin-associated protein, plays a pivotal role in insulin secretion by pancreatic β-cells. It contains a PDZ domain (β2S-PDZ) that, in complex with protein-tyrosine phosphatase ICA512, anchors the dense insulin granules to actin filaments. The phosphorylation state of β2-syntrophin allosterically regulates the affinity of β2S-PDZ for ICA512, and the disruption of the complex triggers the mobilization of the insulin granule stores. Here, we investigate the thermal unfolding of β2S-PDZ at different pH and urea concentrations. Our results indicate that, unlike other PDZ domains, β2S-PDZ is marginally stable. Thermal denaturation experiments show broad transitions and cold denaturation, and a two-state model fit reveals a significant unfolded fraction under physiological conditions. Furthermore, T<sub>m</sub> and T<sub>max</sub> denaturant-dependent shifts and noncoincidence of melting curves monitored at different wavelengths suggest that two-state and three-state models fail to explain the equilibrium data properly and are in better agreement with a downhill scenario. Its higher stability at pH > 9 and the results of molecular dynamics simulations indicate that this behavior of β2S-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 the regulation of insulin secretion.
format Articulo
Articulo
author Torchio, Gabriela María
Ermácora, Mario Roberto
Sica, Mauricio Pablo
author_facet Torchio, Gabriela María
Ermácora, Mario Roberto
Sica, Mauricio Pablo
author_sort Torchio, Gabriela María
title Equilibrium unfolding of the PDZ domain of β2-syntrophin
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
publishDate 2012
url http://sedici.unlp.edu.ar/handle/10915/87841
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AT ermacoramarioroberto equilibriumunfoldingofthepdzdomainofb2syntrophin
AT sicamauriciopablo equilibriumunfoldingofthepdzdomainofb2syntrophin
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