Opposing effects of Na+ and K+ on the thermal stability of Na+,K+-ATPase

Autores
Kaufman, Sergio Benjamín; Gonzalez Flecha, Francisco Luis; Gonzalez-Lebrero, Rodolfo Martin
Año de publicación
2012
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Folding and structural stability are key factors for the proper biological function of proteins. Na+,K+-ATPase is an integral membrane protein involved in the active transport of Na+ and K+ across the plasma membrane. In this work we characterized the effects of K+ and Na+ on the thermal inactivation of Na+,K+-ATPase, evaluating both catalytic and transport capacities of the pump. Both activities of the enzyme decrease with the preincubation time as first-order kinetics. The thermal inactivation of Na+,K+-ATPase is simultaneous with a conformational change detected by tryptophan and 1-aniline-8-naphtalenesulfonate (ANS) fluorescence. The kinetic coefficient of thermal inactivation was affected by the presence of Na+ and K+ (or Rb+) and the temperature of the preincuabtion media. Our results show that K+ or Rb+ stabilize the enzyme, while Na+ decreases the stability of Na+,K+-ATPase. Both effects are exerted by the specific binding of these cations to the pump. Also, we provided strong evidence that the Rb+ (or K+) stabilization effect is due to the occlusion of these cations into the enzyme. Here, we proposed a minimal kinetic model that explains the behavior observed in the experimental results and allows a better understanding of the results presented by other researchers. The thermal inactivation process was also analyzed according to Kramer’s theory.
Fil: Kaufman, Sergio Benjamín. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Biológica; Argentina
Fil: Gonzalez Flecha, Francisco Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Biológica; Argentina
Fil: Gonzalez-Lebrero, Rodolfo Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Biológica; Argentina
Materia
Thermal Inactivation
Na+,K+-Atpase
Conformational Change
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:11336/18230
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/18230
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Opposing effects of Na+ and K+ on the thermal stability of Na+,K+-ATPaseKaufman, Sergio BenjamínGonzalez Flecha, Francisco LuisGonzalez-Lebrero, Rodolfo MartinThermal InactivationNa+,K+-AtpaseConformational Changehttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Folding and structural stability are key factors for the proper biological function of proteins. Na+,K+-ATPase is an integral membrane protein involved in the active transport of Na+ and K+ across the plasma membrane. In this work we characterized the effects of K+ and Na+ on the thermal inactivation of Na+,K+-ATPase, evaluating both catalytic and transport capacities of the pump. Both activities of the enzyme decrease with the preincubation time as first-order kinetics. The thermal inactivation of Na+,K+-ATPase is simultaneous with a conformational change detected by tryptophan and 1-aniline-8-naphtalenesulfonate (ANS) fluorescence. The kinetic coefficient of thermal inactivation was affected by the presence of Na+ and K+ (or Rb+) and the temperature of the preincuabtion media. Our results show that K+ or Rb+ stabilize the enzyme, while Na+ decreases the stability of Na+,K+-ATPase. Both effects are exerted by the specific binding of these cations to the pump. Also, we provided strong evidence that the Rb+ (or K+) stabilization effect is due to the occlusion of these cations into the enzyme. Here, we proposed a minimal kinetic model that explains the behavior observed in the experimental results and allows a better understanding of the results presented by other researchers. The thermal inactivation process was also analyzed according to Kramer’s theory.Fil: Kaufman, Sergio Benjamín. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Biológica; ArgentinaFil: Gonzalez Flecha, Francisco Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Biológica; ArgentinaFil: Gonzalez-Lebrero, Rodolfo Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Biológica; ArgentinaAmerican Chemical Society2012-03info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/18230Kaufman, Sergio Benjamín; Gonzalez Flecha, Francisco Luis; Gonzalez-Lebrero, Rodolfo Martin; Opposing effects of Na+ and K+ on the thermal stability of Na+,K+-ATPase; American Chemical Society; Journal of Physical Chemistry B; 116; 10; 3-2012; 3421-34291520-6106CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1021/jp2124108info:eu-repo/semantics/altIdentifier/url/http://pubs.acs.org/doi/abs/10.1021/jp2124108info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T15:05:32Zoai:ri.conicet.gov.ar:11336/18230instacron: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:34982025-10-15 15:05:32.833CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Opposing effects of Na+ and K+ on the thermal stability of Na+,K+-ATPase
title Opposing effects of Na+ and K+ on the thermal stability of Na+,K+-ATPase
spellingShingle Opposing effects of Na+ and K+ on the thermal stability of Na+,K+-ATPase
Kaufman, Sergio Benjamín
Thermal Inactivation
Na+,K+-Atpase
Conformational Change
title_short Opposing effects of Na+ and K+ on the thermal stability of Na+,K+-ATPase
title_full Opposing effects of Na+ and K+ on the thermal stability of Na+,K+-ATPase
title_fullStr Opposing effects of Na+ and K+ on the thermal stability of Na+,K+-ATPase
title_full_unstemmed Opposing effects of Na+ and K+ on the thermal stability of Na+,K+-ATPase
title_sort Opposing effects of Na+ and K+ on the thermal stability of Na+,K+-ATPase
dc.creator.none.fl_str_mv Kaufman, Sergio Benjamín
Gonzalez Flecha, Francisco Luis
Gonzalez-Lebrero, Rodolfo Martin
author Kaufman, Sergio Benjamín
author_facet Kaufman, Sergio Benjamín
Gonzalez Flecha, Francisco Luis
Gonzalez-Lebrero, Rodolfo Martin
author_role author
author2 Gonzalez Flecha, Francisco Luis
Gonzalez-Lebrero, Rodolfo Martin
author2_role author
author
dc.subject.none.fl_str_mv Thermal Inactivation
Na+,K+-Atpase
Conformational Change
topic Thermal Inactivation
Na+,K+-Atpase
Conformational Change
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Folding and structural stability are key factors for the proper biological function of proteins. Na+,K+-ATPase is an integral membrane protein involved in the active transport of Na+ and K+ across the plasma membrane. In this work we characterized the effects of K+ and Na+ on the thermal inactivation of Na+,K+-ATPase, evaluating both catalytic and transport capacities of the pump. Both activities of the enzyme decrease with the preincubation time as first-order kinetics. The thermal inactivation of Na+,K+-ATPase is simultaneous with a conformational change detected by tryptophan and 1-aniline-8-naphtalenesulfonate (ANS) fluorescence. The kinetic coefficient of thermal inactivation was affected by the presence of Na+ and K+ (or Rb+) and the temperature of the preincuabtion media. Our results show that K+ or Rb+ stabilize the enzyme, while Na+ decreases the stability of Na+,K+-ATPase. Both effects are exerted by the specific binding of these cations to the pump. Also, we provided strong evidence that the Rb+ (or K+) stabilization effect is due to the occlusion of these cations into the enzyme. Here, we proposed a minimal kinetic model that explains the behavior observed in the experimental results and allows a better understanding of the results presented by other researchers. The thermal inactivation process was also analyzed according to Kramer’s theory.
Fil: Kaufman, Sergio Benjamín. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Biológica; Argentina
Fil: Gonzalez Flecha, Francisco Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Biológica; Argentina
Fil: Gonzalez-Lebrero, Rodolfo Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Biológica; Argentina
description Folding and structural stability are key factors for the proper biological function of proteins. Na+,K+-ATPase is an integral membrane protein involved in the active transport of Na+ and K+ across the plasma membrane. In this work we characterized the effects of K+ and Na+ on the thermal inactivation of Na+,K+-ATPase, evaluating both catalytic and transport capacities of the pump. Both activities of the enzyme decrease with the preincubation time as first-order kinetics. The thermal inactivation of Na+,K+-ATPase is simultaneous with a conformational change detected by tryptophan and 1-aniline-8-naphtalenesulfonate (ANS) fluorescence. The kinetic coefficient of thermal inactivation was affected by the presence of Na+ and K+ (or Rb+) and the temperature of the preincuabtion media. Our results show that K+ or Rb+ stabilize the enzyme, while Na+ decreases the stability of Na+,K+-ATPase. Both effects are exerted by the specific binding of these cations to the pump. Also, we provided strong evidence that the Rb+ (or K+) stabilization effect is due to the occlusion of these cations into the enzyme. Here, we proposed a minimal kinetic model that explains the behavior observed in the experimental results and allows a better understanding of the results presented by other researchers. The thermal inactivation process was also analyzed according to Kramer’s theory.
publishDate 2012
dc.date.none.fl_str_mv 2012-03
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/11336/18230
Kaufman, Sergio Benjamín; Gonzalez Flecha, Francisco Luis; Gonzalez-Lebrero, Rodolfo Martin; Opposing effects of Na+ and K+ on the thermal stability of Na+,K+-ATPase; American Chemical Society; Journal of Physical Chemistry B; 116; 10; 3-2012; 3421-3429
1520-6106
CONICET Digital
CONICET
url http://hdl.handle.net/11336/18230
identifier_str_mv Kaufman, Sergio Benjamín; Gonzalez Flecha, Francisco Luis; Gonzalez-Lebrero, Rodolfo Martin; Opposing effects of Na+ and K+ on the thermal stability of Na+,K+-ATPase; American Chemical Society; Journal of Physical Chemistry B; 116; 10; 3-2012; 3421-3429
1520-6106
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1021/jp2124108
info:eu-repo/semantics/altIdentifier/url/http://pubs.acs.org/doi/abs/10.1021/jp2124108
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
application/pdf
dc.publisher.none.fl_str_mv American Chemical Society
publisher.none.fl_str_mv American Chemical Society
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 13.22299

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