The E2P-like state induced by magnesium fluoride complexes in the Na,K-ATPase: kinetics of formation and interaction with Rb+

Autores
Montes, Monica Raquel; Ferreira Gomes, Mariela Soledad; Centeno, Mercedes; Rossi, Rolando Carlos
Año de publicación
2015
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The first X-ray crystal structures of the Na,K-ATPase were obtained in the presence of magnesium and fluoride as E2(K2)Mg–MgF4, an E2∙Pi-like state capable to occlude K+ (or Rb+). This work presents a functional characterization of the crystallized form of the enzyme and proposes a model to explain the interaction between magnesium, fluoride and Rb+ with the Na,K-ATPase. We studied the effect of magnesium and magnesium fluoride complexes on the E1–E2 conformational transition and the kinetics of Rb+ exchange between the medium and the E2(Rb2)Mg–MgF4 state. Our results show that both in the absence and in the presence of Rb+, simultaneous addition of magnesium and fluoride stabilizes the Na,K-ATPase in an E2 conformation, presumably the E2Mg–MgF4 complex, that is unable to shift to E1 upon addition of Na+. The time course of conformational change suggests the action of fluoride and magnesium at different steps of the E2Mg–MgF4 formation. Increasing concentrations of fluoride revert along a sigmoid curve the drop in the level of occluded Rb+ caused by Mg2 +. Na+-induced release of Rb+ from E2(Rb2)Mg–MgF4 occurs at the same rate as from E2(Rb2) but is insensitive to ADP. The rate of Rb+ occlusion into the E2Mg–MgF4 state is 5–8 times lower than that described for the E2Mg–vanadate complex. Since the E2Mg–MgF4 and E2Mg–vanadate complexes represent different intermediates in the E2-P → E2 dephosphorylation sequence, the variation in occlusion rate could provide a tool to discriminate between these intermediates.
Fil: Montes, Monica Raquel. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Biológica; Argentina. 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
Fil: Ferreira Gomes, Mariela Soledad. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Biológica; Argentina. 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
Fil: Centeno, Mercedes. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Biológica; Argentina. 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
Fil: Rossi, Rolando Carlos. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Biológica; Argentina. 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
Materia
P-Type Atpases
Crystallized Na
E2p-Like States
Magnesium Fluoride Complex
Rb+ Occlusion
K-Atpase
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/18326
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/18326
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling The E2P-like state induced by magnesium fluoride complexes in the Na,K-ATPase: kinetics of formation and interaction with Rb+Montes, Monica RaquelFerreira Gomes, Mariela SoledadCenteno, MercedesRossi, Rolando CarlosP-Type AtpasesCrystallized NaE2p-Like StatesMagnesium Fluoride ComplexRb+ OcclusionK-Atpasehttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The first X-ray crystal structures of the Na,K-ATPase were obtained in the presence of magnesium and fluoride as E2(K2)Mg–MgF4, an E2∙Pi-like state capable to occlude K+ (or Rb+). This work presents a functional characterization of the crystallized form of the enzyme and proposes a model to explain the interaction between magnesium, fluoride and Rb+ with the Na,K-ATPase. We studied the effect of magnesium and magnesium fluoride complexes on the E1–E2 conformational transition and the kinetics of Rb+ exchange between the medium and the E2(Rb2)Mg–MgF4 state. Our results show that both in the absence and in the presence of Rb+, simultaneous addition of magnesium and fluoride stabilizes the Na,K-ATPase in an E2 conformation, presumably the E2Mg–MgF4 complex, that is unable to shift to E1 upon addition of Na+. The time course of conformational change suggests the action of fluoride and magnesium at different steps of the E2Mg–MgF4 formation. Increasing concentrations of fluoride revert along a sigmoid curve the drop in the level of occluded Rb+ caused by Mg2 +. Na+-induced release of Rb+ from E2(Rb2)Mg–MgF4 occurs at the same rate as from E2(Rb2) but is insensitive to ADP. The rate of Rb+ occlusion into the E2Mg–MgF4 state is 5–8 times lower than that described for the E2Mg–vanadate complex. Since the E2Mg–MgF4 and E2Mg–vanadate complexes represent different intermediates in the E2-P → E2 dephosphorylation sequence, the variation in occlusion rate could provide a tool to discriminate between these intermediates.Fil: Montes, Monica Raquel. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Biológica; Argentina. 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; ArgentinaFil: Ferreira Gomes, Mariela Soledad. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Biológica; Argentina. 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; ArgentinaFil: Centeno, Mercedes. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Biológica; Argentina. 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; ArgentinaFil: Rossi, Rolando Carlos. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Biológica; Argentina. 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; ArgentinaElsevier Science2015-07info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/18326Montes, Monica Raquel; Ferreira Gomes, Mariela Soledad; Centeno, Mercedes; Rossi, Rolando Carlos; The E2P-like state induced by magnesium fluoride complexes in the Na,K-ATPase: kinetics of formation and interaction with Rb+; Elsevier Science; Biochimica et Biophysica Acta - Biomembranes; 1848; 7; 7-2015; 1514-15230005-2736CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S0005273615001029info:eu-repo/semantics/altIdentifier/doi/10.1016/j.bbamem.2015.03.023info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:38:33Zoai:ri.conicet.gov.ar:11336/18326instacron: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 10:38:33.346CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv The E2P-like state induced by magnesium fluoride complexes in the Na,K-ATPase: kinetics of formation and interaction with Rb+
title The E2P-like state induced by magnesium fluoride complexes in the Na,K-ATPase: kinetics of formation and interaction with Rb+
spellingShingle The E2P-like state induced by magnesium fluoride complexes in the Na,K-ATPase: kinetics of formation and interaction with Rb+
Montes, Monica Raquel
P-Type Atpases
Crystallized Na
E2p-Like States
Magnesium Fluoride Complex
Rb+ Occlusion
K-Atpase
title_short The E2P-like state induced by magnesium fluoride complexes in the Na,K-ATPase: kinetics of formation and interaction with Rb+
title_full The E2P-like state induced by magnesium fluoride complexes in the Na,K-ATPase: kinetics of formation and interaction with Rb+
title_fullStr The E2P-like state induced by magnesium fluoride complexes in the Na,K-ATPase: kinetics of formation and interaction with Rb+
title_full_unstemmed The E2P-like state induced by magnesium fluoride complexes in the Na,K-ATPase: kinetics of formation and interaction with Rb+
title_sort The E2P-like state induced by magnesium fluoride complexes in the Na,K-ATPase: kinetics of formation and interaction with Rb+
dc.creator.none.fl_str_mv Montes, Monica Raquel
Ferreira Gomes, Mariela Soledad
Centeno, Mercedes
Rossi, Rolando Carlos
author Montes, Monica Raquel
author_facet Montes, Monica Raquel
Ferreira Gomes, Mariela Soledad
Centeno, Mercedes
Rossi, Rolando Carlos
author_role author
author2 Ferreira Gomes, Mariela Soledad
Centeno, Mercedes
Rossi, Rolando Carlos
author2_role author
author
author
dc.subject.none.fl_str_mv P-Type Atpases
Crystallized Na
E2p-Like States
Magnesium Fluoride Complex
Rb+ Occlusion
K-Atpase
topic P-Type Atpases
Crystallized Na
E2p-Like States
Magnesium Fluoride Complex
Rb+ Occlusion
K-Atpase
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv The first X-ray crystal structures of the Na,K-ATPase were obtained in the presence of magnesium and fluoride as E2(K2)Mg–MgF4, an E2∙Pi-like state capable to occlude K+ (or Rb+). This work presents a functional characterization of the crystallized form of the enzyme and proposes a model to explain the interaction between magnesium, fluoride and Rb+ with the Na,K-ATPase. We studied the effect of magnesium and magnesium fluoride complexes on the E1–E2 conformational transition and the kinetics of Rb+ exchange between the medium and the E2(Rb2)Mg–MgF4 state. Our results show that both in the absence and in the presence of Rb+, simultaneous addition of magnesium and fluoride stabilizes the Na,K-ATPase in an E2 conformation, presumably the E2Mg–MgF4 complex, that is unable to shift to E1 upon addition of Na+. The time course of conformational change suggests the action of fluoride and magnesium at different steps of the E2Mg–MgF4 formation. Increasing concentrations of fluoride revert along a sigmoid curve the drop in the level of occluded Rb+ caused by Mg2 +. Na+-induced release of Rb+ from E2(Rb2)Mg–MgF4 occurs at the same rate as from E2(Rb2) but is insensitive to ADP. The rate of Rb+ occlusion into the E2Mg–MgF4 state is 5–8 times lower than that described for the E2Mg–vanadate complex. Since the E2Mg–MgF4 and E2Mg–vanadate complexes represent different intermediates in the E2-P → E2 dephosphorylation sequence, the variation in occlusion rate could provide a tool to discriminate between these intermediates.
Fil: Montes, Monica Raquel. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Biológica; Argentina. 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
Fil: Ferreira Gomes, Mariela Soledad. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Biológica; Argentina. 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
Fil: Centeno, Mercedes. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Biológica; Argentina. 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
Fil: Rossi, Rolando Carlos. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Biológica; Argentina. 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
description The first X-ray crystal structures of the Na,K-ATPase were obtained in the presence of magnesium and fluoride as E2(K2)Mg–MgF4, an E2∙Pi-like state capable to occlude K+ (or Rb+). This work presents a functional characterization of the crystallized form of the enzyme and proposes a model to explain the interaction between magnesium, fluoride and Rb+ with the Na,K-ATPase. We studied the effect of magnesium and magnesium fluoride complexes on the E1–E2 conformational transition and the kinetics of Rb+ exchange between the medium and the E2(Rb2)Mg–MgF4 state. Our results show that both in the absence and in the presence of Rb+, simultaneous addition of magnesium and fluoride stabilizes the Na,K-ATPase in an E2 conformation, presumably the E2Mg–MgF4 complex, that is unable to shift to E1 upon addition of Na+. The time course of conformational change suggests the action of fluoride and magnesium at different steps of the E2Mg–MgF4 formation. Increasing concentrations of fluoride revert along a sigmoid curve the drop in the level of occluded Rb+ caused by Mg2 +. Na+-induced release of Rb+ from E2(Rb2)Mg–MgF4 occurs at the same rate as from E2(Rb2) but is insensitive to ADP. The rate of Rb+ occlusion into the E2Mg–MgF4 state is 5–8 times lower than that described for the E2Mg–vanadate complex. Since the E2Mg–MgF4 and E2Mg–vanadate complexes represent different intermediates in the E2-P → E2 dephosphorylation sequence, the variation in occlusion rate could provide a tool to discriminate between these intermediates.
publishDate 2015
dc.date.none.fl_str_mv 2015-07
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/11336/18326
Montes, Monica Raquel; Ferreira Gomes, Mariela Soledad; Centeno, Mercedes; Rossi, Rolando Carlos; The E2P-like state induced by magnesium fluoride complexes in the Na,K-ATPase: kinetics of formation and interaction with Rb+; Elsevier Science; Biochimica et Biophysica Acta - Biomembranes; 1848; 7; 7-2015; 1514-1523
0005-2736
CONICET Digital
CONICET
url http://hdl.handle.net/11336/18326
identifier_str_mv Montes, Monica Raquel; Ferreira Gomes, Mariela Soledad; Centeno, Mercedes; Rossi, Rolando Carlos; The E2P-like state induced by magnesium fluoride complexes in the Na,K-ATPase: kinetics of formation and interaction with Rb+; Elsevier Science; Biochimica et Biophysica Acta - Biomembranes; 1848; 7; 7-2015; 1514-1523
0005-2736
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://www.sciencedirect.com/science/article/pii/S0005273615001029
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.bbamem.2015.03.023
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier Science
publisher.none.fl_str_mv Elsevier Science
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 13.070432

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