Metabolic regulation of the squid nerve Na+/Ca2+ exchanger: Recent kinetic, biochemical and structural developments

Autores
Elso, Graciela Raquel; Podjarny, Alberto Daniel; DiPolo, Reinaldo; Beauge, Luis Alberto
Año de publicación
2012
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The Naþ/Ca2þ exchangers are structural membrane proteins, essential for the extrusion of Ca2þ from most animal cells. Apart from the transport sites, they have several interacting ionic and metabolic sites located at the intracellular loop of the exchanger protein. One of these, the intracellular Ca2þ regulatory sites, are essential and must be occupied by Ca2þ to allow any type of ion (Naþ or Ca2þ) translocation. Intracellular protons and Naþ are inhibitory by reducing the affinity of the regulatory sites for Ca2þ; MgATP stimulates by antagonizing Hþ and Naþ. We have proposed a kinetic scheme to explain all ionic and metabolic regulation of the squid nerve Naþ/Ca2þ exchanger. This model uniquely accounts for most of the new kinetic data provided here; however, none of the existing models can explain the trans effects of the Cai 2þ-regulatory sites on external cation transport sites; i.e. all models are incomplete. MgATP up-regulation of the squid Naþ/Ca2þ exchanger requires a cytosolic protein, which has been recently identified as a member of the lipocalin super family of Lipid Binding Proteins (LBP or FABP) of 132 amino acids (ReP1-NCXSQ, access to GenBank EU981897). This protein was cloned, expressed and purified. To be active, ReP1-NCXSQ must be phosphorylated from MgATP by a kinase present in the plasma membrane. Phosphorylated ReP1-NCXSQ can stimulate the exchanger in the absence of ATP. Experiments with proteoliposomes proved that this up-regulation can take place just with the lipid membrane and the exchanger protein. The structure of ReP1-NCXSQ predicted from the amino acid sequence has been confirmed by X-ray crystal analysis; it has a “barrel” formed by ten beta sheets and two alpha helices, with a lipid coordinated by hydrogen bonds with Arg 126 and Tyr 128.
Fil: Elso, Graciela Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; Argentina
Fil: Podjarny, Alberto Daniel. Centre National de la Recherche Scientifique. Igbmc; Francia. Université de Strasbourg; Francia
Fil: DiPolo, Reinaldo. Instituto Venezolano de Investigaciones Científicas; Venezuela
Fil: Beauge, Luis Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; Argentina
Materia
Na+/Ca2+exchanger
Squid nerve
Metabolic regulation
Regulatory protein
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:11336/268610
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/268610
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Metabolic regulation of the squid nerve Na+/Ca2+ exchanger: Recent kinetic, biochemical and structural developmentsElso, Graciela RaquelPodjarny, Alberto DanielDiPolo, ReinaldoBeauge, Luis AlbertoNa+/Ca2+exchangerSquid nerveMetabolic regulationRegulatory proteinhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The Naþ/Ca2þ exchangers are structural membrane proteins, essential for the extrusion of Ca2þ from most animal cells. Apart from the transport sites, they have several interacting ionic and metabolic sites located at the intracellular loop of the exchanger protein. One of these, the intracellular Ca2þ regulatory sites, are essential and must be occupied by Ca2þ to allow any type of ion (Naþ or Ca2þ) translocation. Intracellular protons and Naþ are inhibitory by reducing the affinity of the regulatory sites for Ca2þ; MgATP stimulates by antagonizing Hþ and Naþ. We have proposed a kinetic scheme to explain all ionic and metabolic regulation of the squid nerve Naþ/Ca2þ exchanger. This model uniquely accounts for most of the new kinetic data provided here; however, none of the existing models can explain the trans effects of the Cai 2þ-regulatory sites on external cation transport sites; i.e. all models are incomplete. MgATP up-regulation of the squid Naþ/Ca2þ exchanger requires a cytosolic protein, which has been recently identified as a member of the lipocalin super family of Lipid Binding Proteins (LBP or FABP) of 132 amino acids (ReP1-NCXSQ, access to GenBank EU981897). This protein was cloned, expressed and purified. To be active, ReP1-NCXSQ must be phosphorylated from MgATP by a kinase present in the plasma membrane. Phosphorylated ReP1-NCXSQ can stimulate the exchanger in the absence of ATP. Experiments with proteoliposomes proved that this up-regulation can take place just with the lipid membrane and the exchanger protein. The structure of ReP1-NCXSQ predicted from the amino acid sequence has been confirmed by X-ray crystal analysis; it has a “barrel” formed by ten beta sheets and two alpha helices, with a lipid coordinated by hydrogen bonds with Arg 126 and Tyr 128.Fil: Elso, Graciela Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; ArgentinaFil: Podjarny, Alberto Daniel. Centre National de la Recherche Scientifique. Igbmc; Francia. Université de Strasbourg; FranciaFil: DiPolo, Reinaldo. Instituto Venezolano de Investigaciones Científicas; VenezuelaFil: Beauge, Luis Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; ArgentinaPergamon-Elsevier Science Ltd2012-01info: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/11336/268610Elso, Graciela Raquel; Podjarny, Alberto Daniel; DiPolo, Reinaldo; Beauge, Luis Alberto; Metabolic regulation of the squid nerve Na+/Ca2+ exchanger: Recent kinetic, biochemical and structural developments; Pergamon-Elsevier Science Ltd; Progress In Biophysics And Molecular Biology; 108; 1-2; 1-2012; 47-630079-6107CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/abs/pii/S0079610711001052info:eu-repo/semantics/altIdentifier/doi/10.1016/j.pbiomolbio.2011.09.006info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:32:28Zoai:ri.conicet.gov.ar:11336/268610instacron: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 10:32:28.691CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Metabolic regulation of the squid nerve Na+/Ca2+ exchanger: Recent kinetic, biochemical and structural developments
title Metabolic regulation of the squid nerve Na+/Ca2+ exchanger: Recent kinetic, biochemical and structural developments
spellingShingle Metabolic regulation of the squid nerve Na+/Ca2+ exchanger: Recent kinetic, biochemical and structural developments
Elso, Graciela Raquel
Na+/Ca2+exchanger
Squid nerve
Metabolic regulation
Regulatory protein
title_short Metabolic regulation of the squid nerve Na+/Ca2+ exchanger: Recent kinetic, biochemical and structural developments
title_full Metabolic regulation of the squid nerve Na+/Ca2+ exchanger: Recent kinetic, biochemical and structural developments
title_fullStr Metabolic regulation of the squid nerve Na+/Ca2+ exchanger: Recent kinetic, biochemical and structural developments
title_full_unstemmed Metabolic regulation of the squid nerve Na+/Ca2+ exchanger: Recent kinetic, biochemical and structural developments
title_sort Metabolic regulation of the squid nerve Na+/Ca2+ exchanger: Recent kinetic, biochemical and structural developments
dc.creator.none.fl_str_mv Elso, Graciela Raquel
Podjarny, Alberto Daniel
DiPolo, Reinaldo
Beauge, Luis Alberto
author Elso, Graciela Raquel
author_facet Elso, Graciela Raquel
Podjarny, Alberto Daniel
DiPolo, Reinaldo
Beauge, Luis Alberto
author_role author
author2 Podjarny, Alberto Daniel
DiPolo, Reinaldo
Beauge, Luis Alberto
author2_role author
author
author
dc.subject.none.fl_str_mv Na+/Ca2+exchanger
Squid nerve
Metabolic regulation
Regulatory protein
topic Na+/Ca2+exchanger
Squid nerve
Metabolic regulation
Regulatory protein
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 Naþ/Ca2þ exchangers are structural membrane proteins, essential for the extrusion of Ca2þ from most animal cells. Apart from the transport sites, they have several interacting ionic and metabolic sites located at the intracellular loop of the exchanger protein. One of these, the intracellular Ca2þ regulatory sites, are essential and must be occupied by Ca2þ to allow any type of ion (Naþ or Ca2þ) translocation. Intracellular protons and Naþ are inhibitory by reducing the affinity of the regulatory sites for Ca2þ; MgATP stimulates by antagonizing Hþ and Naþ. We have proposed a kinetic scheme to explain all ionic and metabolic regulation of the squid nerve Naþ/Ca2þ exchanger. This model uniquely accounts for most of the new kinetic data provided here; however, none of the existing models can explain the trans effects of the Cai 2þ-regulatory sites on external cation transport sites; i.e. all models are incomplete. MgATP up-regulation of the squid Naþ/Ca2þ exchanger requires a cytosolic protein, which has been recently identified as a member of the lipocalin super family of Lipid Binding Proteins (LBP or FABP) of 132 amino acids (ReP1-NCXSQ, access to GenBank EU981897). This protein was cloned, expressed and purified. To be active, ReP1-NCXSQ must be phosphorylated from MgATP by a kinase present in the plasma membrane. Phosphorylated ReP1-NCXSQ can stimulate the exchanger in the absence of ATP. Experiments with proteoliposomes proved that this up-regulation can take place just with the lipid membrane and the exchanger protein. The structure of ReP1-NCXSQ predicted from the amino acid sequence has been confirmed by X-ray crystal analysis; it has a “barrel” formed by ten beta sheets and two alpha helices, with a lipid coordinated by hydrogen bonds with Arg 126 and Tyr 128.
Fil: Elso, Graciela Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; Argentina
Fil: Podjarny, Alberto Daniel. Centre National de la Recherche Scientifique. Igbmc; Francia. Université de Strasbourg; Francia
Fil: DiPolo, Reinaldo. Instituto Venezolano de Investigaciones Científicas; Venezuela
Fil: Beauge, Luis Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; Argentina
description The Naþ/Ca2þ exchangers are structural membrane proteins, essential for the extrusion of Ca2þ from most animal cells. Apart from the transport sites, they have several interacting ionic and metabolic sites located at the intracellular loop of the exchanger protein. One of these, the intracellular Ca2þ regulatory sites, are essential and must be occupied by Ca2þ to allow any type of ion (Naþ or Ca2þ) translocation. Intracellular protons and Naþ are inhibitory by reducing the affinity of the regulatory sites for Ca2þ; MgATP stimulates by antagonizing Hþ and Naþ. We have proposed a kinetic scheme to explain all ionic and metabolic regulation of the squid nerve Naþ/Ca2þ exchanger. This model uniquely accounts for most of the new kinetic data provided here; however, none of the existing models can explain the trans effects of the Cai 2þ-regulatory sites on external cation transport sites; i.e. all models are incomplete. MgATP up-regulation of the squid Naþ/Ca2þ exchanger requires a cytosolic protein, which has been recently identified as a member of the lipocalin super family of Lipid Binding Proteins (LBP or FABP) of 132 amino acids (ReP1-NCXSQ, access to GenBank EU981897). This protein was cloned, expressed and purified. To be active, ReP1-NCXSQ must be phosphorylated from MgATP by a kinase present in the plasma membrane. Phosphorylated ReP1-NCXSQ can stimulate the exchanger in the absence of ATP. Experiments with proteoliposomes proved that this up-regulation can take place just with the lipid membrane and the exchanger protein. The structure of ReP1-NCXSQ predicted from the amino acid sequence has been confirmed by X-ray crystal analysis; it has a “barrel” formed by ten beta sheets and two alpha helices, with a lipid coordinated by hydrogen bonds with Arg 126 and Tyr 128.
publishDate 2012
dc.date.none.fl_str_mv 2012-01
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/11336/268610
Elso, Graciela Raquel; Podjarny, Alberto Daniel; DiPolo, Reinaldo; Beauge, Luis Alberto; Metabolic regulation of the squid nerve Na+/Ca2+ exchanger: Recent kinetic, biochemical and structural developments; Pergamon-Elsevier Science Ltd; Progress In Biophysics And Molecular Biology; 108; 1-2; 1-2012; 47-63
0079-6107
CONICET Digital
CONICET
url http://hdl.handle.net/11336/268610
identifier_str_mv Elso, Graciela Raquel; Podjarny, Alberto Daniel; DiPolo, Reinaldo; Beauge, Luis Alberto; Metabolic regulation of the squid nerve Na+/Ca2+ exchanger: Recent kinetic, biochemical and structural developments; Pergamon-Elsevier Science Ltd; Progress In Biophysics And Molecular Biology; 108; 1-2; 1-2012; 47-63
0079-6107
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/abs/pii/S0079610711001052
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.pbiomolbio.2011.09.006
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
application/pdf
dc.publisher.none.fl_str_mv Pergamon-Elsevier Science Ltd
publisher.none.fl_str_mv Pergamon-Elsevier Science Ltd
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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