Unexpected Effects of K + and Adenosine Triphosphate on the Thermal Stability of Na + ,K + -ATPase

Autores
Placenti, Maria Agueda; Kaufman, Sergio Benjamín; Gonzalez Flecha, Francisco Luis; Gonzalez-Lebrero, Rodolfo Martin
Año de publicación
2017
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Na+,K+-ATPase is an integral membrane protein which couples ATP hydrolysis to the transport of three Na+ out and two K+ into the cell. The aim of this work is to characterize the effect of K+, ATP, and Mg2+ (essential activator) on the Na+,K+-ATPase thermal stability. Under all conditions tested, thermal inactivation of the enzyme is concomitant with a structural change involving the ATP binding site and membrane-associated regions. Both ligands exert a clear stabilizing effect due to both enthalpic and entropic contributions. Competition experiments between ATP and K+ showed that, when ATP is present, the inactivation rate coefficient exhibits a biphasic dependence on K+ concentration. At low [K+], destabilization of the enzyme is observed, while stabilization occurred at larger cation concentrations. This is not expected for a simple competition between the enzyme and two ligands that individually protect the enzyme. A model that includes enzyme species with none, one, or two K+ and/or one molecule of ATP bound explains the experimental data. We concluded that, despite both ligands stabilizing the enzyme, the species with one K+ and one ATP simultaneously bound is unstable.
Fil: Placenti, Maria Agueda. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas ; Argentina
Fil: Kaufman, Sergio Benjamín. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas ; 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 ; 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 ; Argentina
Materia
Na+,K+-ATPase
Membrane protein
Transition State Theory
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/47222
Acceder
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network_name_str CONICET Digital (CONICET)
spelling Unexpected Effects of K + and Adenosine Triphosphate on the Thermal Stability of Na + ,K + -ATPasePlacenti, Maria AguedaKaufman, Sergio BenjamínGonzalez Flecha, Francisco LuisGonzalez-Lebrero, Rodolfo MartinNa+,K+-ATPaseMembrane proteinTransition State Theoryhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Na+,K+-ATPase is an integral membrane protein which couples ATP hydrolysis to the transport of three Na+ out and two K+ into the cell. The aim of this work is to characterize the effect of K+, ATP, and Mg2+ (essential activator) on the Na+,K+-ATPase thermal stability. Under all conditions tested, thermal inactivation of the enzyme is concomitant with a structural change involving the ATP binding site and membrane-associated regions. Both ligands exert a clear stabilizing effect due to both enthalpic and entropic contributions. Competition experiments between ATP and K+ showed that, when ATP is present, the inactivation rate coefficient exhibits a biphasic dependence on K+ concentration. At low [K+], destabilization of the enzyme is observed, while stabilization occurred at larger cation concentrations. This is not expected for a simple competition between the enzyme and two ligands that individually protect the enzyme. A model that includes enzyme species with none, one, or two K+ and/or one molecule of ATP bound explains the experimental data. We concluded that, despite both ligands stabilizing the enzyme, the species with one K+ and one ATP simultaneously bound is unstable.Fil: Placenti, Maria Agueda. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas ; ArgentinaFil: Kaufman, Sergio Benjamín. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas ; ArgentinaFil: Gonzalez Flecha, Francisco Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas ; ArgentinaFil: Gonzalez-Lebrero, Rodolfo Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas ; ArgentinaAmerican Chemical Society2017-05info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/47222Placenti, Maria Agueda; Kaufman, Sergio Benjamín; Gonzalez Flecha, Francisco Luis; Gonzalez-Lebrero, Rodolfo Martin; Unexpected Effects of K + and Adenosine Triphosphate on the Thermal Stability of Na + ,K + -ATPase; American Chemical Society; Journal of Physical Chemistry B; 121; 19; 5-2017; 4949-49571520-6106CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://pubs.acs.org/doi/abs/10.1021/acs.jpcb.7b00629info:eu-repo/semantics/altIdentifier/doi/10.1021/acs.jpcb.7b00629info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-22T12:10:15Zoai:ri.conicet.gov.ar:11336/47222instacron: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:34982025-10-22 12:10:16.103CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Unexpected Effects of K + and Adenosine Triphosphate on the Thermal Stability of Na + ,K + -ATPase
title Unexpected Effects of K + and Adenosine Triphosphate on the Thermal Stability of Na + ,K + -ATPase
spellingShingle Unexpected Effects of K + and Adenosine Triphosphate on the Thermal Stability of Na + ,K + -ATPase
Placenti, Maria Agueda
Na+,K+-ATPase
Membrane protein
Transition State Theory
title_short Unexpected Effects of K + and Adenosine Triphosphate on the Thermal Stability of Na + ,K + -ATPase
title_full Unexpected Effects of K + and Adenosine Triphosphate on the Thermal Stability of Na + ,K + -ATPase
title_fullStr Unexpected Effects of K + and Adenosine Triphosphate on the Thermal Stability of Na + ,K + -ATPase
title_full_unstemmed Unexpected Effects of K + and Adenosine Triphosphate on the Thermal Stability of Na + ,K + -ATPase
title_sort Unexpected Effects of K + and Adenosine Triphosphate on the Thermal Stability of Na + ,K + -ATPase
dc.creator.none.fl_str_mv Placenti, Maria Agueda
Kaufman, Sergio Benjamín
Gonzalez Flecha, Francisco Luis
Gonzalez-Lebrero, Rodolfo Martin
author Placenti, Maria Agueda
author_facet Placenti, Maria Agueda
Kaufman, Sergio Benjamín
Gonzalez Flecha, Francisco Luis
Gonzalez-Lebrero, Rodolfo Martin
author_role author
author2 Kaufman, Sergio Benjamín
Gonzalez Flecha, Francisco Luis
Gonzalez-Lebrero, Rodolfo Martin
author2_role author
author
author
dc.subject.none.fl_str_mv Na+,K+-ATPase
Membrane protein
Transition State Theory
topic Na+,K+-ATPase
Membrane protein
Transition State Theory
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Na+,K+-ATPase is an integral membrane protein which couples ATP hydrolysis to the transport of three Na+ out and two K+ into the cell. The aim of this work is to characterize the effect of K+, ATP, and Mg2+ (essential activator) on the Na+,K+-ATPase thermal stability. Under all conditions tested, thermal inactivation of the enzyme is concomitant with a structural change involving the ATP binding site and membrane-associated regions. Both ligands exert a clear stabilizing effect due to both enthalpic and entropic contributions. Competition experiments between ATP and K+ showed that, when ATP is present, the inactivation rate coefficient exhibits a biphasic dependence on K+ concentration. At low [K+], destabilization of the enzyme is observed, while stabilization occurred at larger cation concentrations. This is not expected for a simple competition between the enzyme and two ligands that individually protect the enzyme. A model that includes enzyme species with none, one, or two K+ and/or one molecule of ATP bound explains the experimental data. We concluded that, despite both ligands stabilizing the enzyme, the species with one K+ and one ATP simultaneously bound is unstable.
Fil: Placenti, Maria Agueda. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas ; Argentina
Fil: Kaufman, Sergio Benjamín. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas ; 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 ; 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 ; Argentina
description Na+,K+-ATPase is an integral membrane protein which couples ATP hydrolysis to the transport of three Na+ out and two K+ into the cell. The aim of this work is to characterize the effect of K+, ATP, and Mg2+ (essential activator) on the Na+,K+-ATPase thermal stability. Under all conditions tested, thermal inactivation of the enzyme is concomitant with a structural change involving the ATP binding site and membrane-associated regions. Both ligands exert a clear stabilizing effect due to both enthalpic and entropic contributions. Competition experiments between ATP and K+ showed that, when ATP is present, the inactivation rate coefficient exhibits a biphasic dependence on K+ concentration. At low [K+], destabilization of the enzyme is observed, while stabilization occurred at larger cation concentrations. This is not expected for a simple competition between the enzyme and two ligands that individually protect the enzyme. A model that includes enzyme species with none, one, or two K+ and/or one molecule of ATP bound explains the experimental data. We concluded that, despite both ligands stabilizing the enzyme, the species with one K+ and one ATP simultaneously bound is unstable.
publishDate 2017
dc.date.none.fl_str_mv 2017-05
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/11336/47222
Placenti, Maria Agueda; Kaufman, Sergio Benjamín; Gonzalez Flecha, Francisco Luis; Gonzalez-Lebrero, Rodolfo Martin; Unexpected Effects of K + and Adenosine Triphosphate on the Thermal Stability of Na + ,K + -ATPase; American Chemical Society; Journal of Physical Chemistry B; 121; 19; 5-2017; 4949-4957
1520-6106
CONICET Digital
CONICET
url http://hdl.handle.net/11336/47222
identifier_str_mv Placenti, Maria Agueda; Kaufman, Sergio Benjamín; Gonzalez Flecha, Francisco Luis; Gonzalez-Lebrero, Rodolfo Martin; Unexpected Effects of K + and Adenosine Triphosphate on the Thermal Stability of Na + ,K + -ATPase; American Chemical Society; Journal of Physical Chemistry B; 121; 19; 5-2017; 4949-4957
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/url/http://pubs.acs.org/doi/abs/10.1021/acs.jpcb.7b00629
info:eu-repo/semantics/altIdentifier/doi/10.1021/acs.jpcb.7b00629
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv 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
_version_ 1846782486550413312
score 12.982451

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