Plasma Membrane Calcium Pump (PMCA) Differential Exposure of Hydrophobic Domains after Calmodulin and Phosphatidic Acid Activation

Autores
Mangialavori, Irene Cecilia; Villamil Giraldo, Ana María; Pignataro, María Florencia; Ferreira Gomes, Mariela Soledad; Caride, Ariel J.; Rossi, Juan Pablo Francisco
Año de publicación
2011
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The exposure of the plasma membrane calcium pump (PMCA) to the surrounding phospholipids was assessed by measuring the incorporation of the photoactivatable phosphatidylcholine analog [125I]TID-PC/16 to the protein. In the presence of Ca2+ both calmodulin (CaM) and phosphatidic acid (PA) greatly decreased the incorporation of [125I]TID-PC/16 to PMCA. Proteolysis of PMCA with V8 protease results in three main fragments: N, which includes transmembrane segments M1 and M2; M, which includes M3 and M4; and C, which includes M5 to M10. CaM decreased the level of incorporation of [125I]TID-PC/16 to fragments M and C, whereas phosphatidic acid decreased the incorporation of [125I]TID-PC/16 to fragments N and M. This suggests that the conformational changes induced by binding of CaM or PA extend to the adjacent transmembrane domains. Interestingly, this result also denotes differences between the active conformations produced by CaM and PA. To verify this point, we measured resonance energy transfer between PMCA labeled with eosin isothiocyanate at the ATP-binding site and the phospholipid RhoPE included in PMCA micelles. CaM decreased the efficiency of the energy transfer between these two probes, whereas PA did not. This result indicates that activation by CaM increases the distance between the ATP-binding site and the membrane, but PA does not affect this distance. Our results disclose main differences between PMCA conformations induced by CaM or PA and show that those differences involve transmembrane regions.
Fil: Mangialavori, Irene Cecilia. 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: Villamil Giraldo, Ana María. 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: Pignataro, María Florencia. 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. 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: Caride, Ariel J.. Mayo Clinic College of Medicine; Estados Unidos
Fil: Rossi, Juan Pablo Francisco. 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
Calcium Atpase
Calmodulin
Phosphatidic Acid
Photoaffinity Labeling
Plasma Membrane
Transmembrane Domain
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:11336/18237
Acceder
id CONICETDig_25d5a337ec8e34e7d905e4cd792bc6c8
oai_identifier_str oai:ri.conicet.gov.ar:11336/18237
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Plasma Membrane Calcium Pump (PMCA) Differential Exposure of Hydrophobic Domains after Calmodulin and Phosphatidic Acid ActivationMangialavori, Irene CeciliaVillamil Giraldo, Ana MaríaPignataro, María FlorenciaFerreira Gomes, Mariela SoledadCaride, Ariel J.Rossi, Juan Pablo FranciscoCalcium AtpaseCalmodulinPhosphatidic AcidPhotoaffinity LabelingPlasma MembraneTransmembrane Domainhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The exposure of the plasma membrane calcium pump (PMCA) to the surrounding phospholipids was assessed by measuring the incorporation of the photoactivatable phosphatidylcholine analog [125I]TID-PC/16 to the protein. In the presence of Ca2+ both calmodulin (CaM) and phosphatidic acid (PA) greatly decreased the incorporation of [125I]TID-PC/16 to PMCA. Proteolysis of PMCA with V8 protease results in three main fragments: N, which includes transmembrane segments M1 and M2; M, which includes M3 and M4; and C, which includes M5 to M10. CaM decreased the level of incorporation of [125I]TID-PC/16 to fragments M and C, whereas phosphatidic acid decreased the incorporation of [125I]TID-PC/16 to fragments N and M. This suggests that the conformational changes induced by binding of CaM or PA extend to the adjacent transmembrane domains. Interestingly, this result also denotes differences between the active conformations produced by CaM and PA. To verify this point, we measured resonance energy transfer between PMCA labeled with eosin isothiocyanate at the ATP-binding site and the phospholipid RhoPE included in PMCA micelles. CaM decreased the efficiency of the energy transfer between these two probes, whereas PA did not. This result indicates that activation by CaM increases the distance between the ATP-binding site and the membrane, but PA does not affect this distance. Our results disclose main differences between PMCA conformations induced by CaM or PA and show that those differences involve transmembrane regions.Fil: Mangialavori, Irene Cecilia. 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: Villamil Giraldo, Ana María. 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: Pignataro, María Florencia. 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. 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: Caride, Ariel J.. Mayo Clinic College of Medicine; Estados UnidosFil: Rossi, Juan Pablo Francisco. 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; ArgentinaAmerican Society for Biochemistry and Molecular Biology2011-05info: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/11336/18237Mangialavori, Irene Cecilia; Villamil Giraldo, Ana María; Pignataro, María Florencia; Ferreira Gomes, Mariela Soledad; Caride, Ariel J.; et al.; Plasma Membrane Calcium Pump (PMCA) Differential Exposure of Hydrophobic Domains after Calmodulin and Phosphatidic Acid Activation; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry; 286; 21; 5-2011; 18397-184040021-92581083-351XCONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0021925820510971info:eu-repo/semantics/altIdentifier/doi/10.1074/jbc.M110.210088info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:08:29Zoai:ri.conicet.gov.ar:11336/18237instacron: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:34982025-09-03 10:08:29.845CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Plasma Membrane Calcium Pump (PMCA) Differential Exposure of Hydrophobic Domains after Calmodulin and Phosphatidic Acid Activation
title Plasma Membrane Calcium Pump (PMCA) Differential Exposure of Hydrophobic Domains after Calmodulin and Phosphatidic Acid Activation
spellingShingle Plasma Membrane Calcium Pump (PMCA) Differential Exposure of Hydrophobic Domains after Calmodulin and Phosphatidic Acid Activation
Mangialavori, Irene Cecilia
Calcium Atpase
Calmodulin
Phosphatidic Acid
Photoaffinity Labeling
Plasma Membrane
Transmembrane Domain
title_short Plasma Membrane Calcium Pump (PMCA) Differential Exposure of Hydrophobic Domains after Calmodulin and Phosphatidic Acid Activation
title_full Plasma Membrane Calcium Pump (PMCA) Differential Exposure of Hydrophobic Domains after Calmodulin and Phosphatidic Acid Activation
title_fullStr Plasma Membrane Calcium Pump (PMCA) Differential Exposure of Hydrophobic Domains after Calmodulin and Phosphatidic Acid Activation
title_full_unstemmed Plasma Membrane Calcium Pump (PMCA) Differential Exposure of Hydrophobic Domains after Calmodulin and Phosphatidic Acid Activation
title_sort Plasma Membrane Calcium Pump (PMCA) Differential Exposure of Hydrophobic Domains after Calmodulin and Phosphatidic Acid Activation
dc.creator.none.fl_str_mv Mangialavori, Irene Cecilia
Villamil Giraldo, Ana María
Pignataro, María Florencia
Ferreira Gomes, Mariela Soledad
Caride, Ariel J.
Rossi, Juan Pablo Francisco
author Mangialavori, Irene Cecilia
author_facet Mangialavori, Irene Cecilia
Villamil Giraldo, Ana María
Pignataro, María Florencia
Ferreira Gomes, Mariela Soledad
Caride, Ariel J.
Rossi, Juan Pablo Francisco
author_role author
author2 Villamil Giraldo, Ana María
Pignataro, María Florencia
Ferreira Gomes, Mariela Soledad
Caride, Ariel J.
Rossi, Juan Pablo Francisco
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Calcium Atpase
Calmodulin
Phosphatidic Acid
Photoaffinity Labeling
Plasma Membrane
Transmembrane Domain
topic Calcium Atpase
Calmodulin
Phosphatidic Acid
Photoaffinity Labeling
Plasma Membrane
Transmembrane Domain
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 exposure of the plasma membrane calcium pump (PMCA) to the surrounding phospholipids was assessed by measuring the incorporation of the photoactivatable phosphatidylcholine analog [125I]TID-PC/16 to the protein. In the presence of Ca2+ both calmodulin (CaM) and phosphatidic acid (PA) greatly decreased the incorporation of [125I]TID-PC/16 to PMCA. Proteolysis of PMCA with V8 protease results in three main fragments: N, which includes transmembrane segments M1 and M2; M, which includes M3 and M4; and C, which includes M5 to M10. CaM decreased the level of incorporation of [125I]TID-PC/16 to fragments M and C, whereas phosphatidic acid decreased the incorporation of [125I]TID-PC/16 to fragments N and M. This suggests that the conformational changes induced by binding of CaM or PA extend to the adjacent transmembrane domains. Interestingly, this result also denotes differences between the active conformations produced by CaM and PA. To verify this point, we measured resonance energy transfer between PMCA labeled with eosin isothiocyanate at the ATP-binding site and the phospholipid RhoPE included in PMCA micelles. CaM decreased the efficiency of the energy transfer between these two probes, whereas PA did not. This result indicates that activation by CaM increases the distance between the ATP-binding site and the membrane, but PA does not affect this distance. Our results disclose main differences between PMCA conformations induced by CaM or PA and show that those differences involve transmembrane regions.
Fil: Mangialavori, Irene Cecilia. 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: Villamil Giraldo, Ana María. 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: Pignataro, María Florencia. 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. 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: Caride, Ariel J.. Mayo Clinic College of Medicine; Estados Unidos
Fil: Rossi, Juan Pablo Francisco. 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 exposure of the plasma membrane calcium pump (PMCA) to the surrounding phospholipids was assessed by measuring the incorporation of the photoactivatable phosphatidylcholine analog [125I]TID-PC/16 to the protein. In the presence of Ca2+ both calmodulin (CaM) and phosphatidic acid (PA) greatly decreased the incorporation of [125I]TID-PC/16 to PMCA. Proteolysis of PMCA with V8 protease results in three main fragments: N, which includes transmembrane segments M1 and M2; M, which includes M3 and M4; and C, which includes M5 to M10. CaM decreased the level of incorporation of [125I]TID-PC/16 to fragments M and C, whereas phosphatidic acid decreased the incorporation of [125I]TID-PC/16 to fragments N and M. This suggests that the conformational changes induced by binding of CaM or PA extend to the adjacent transmembrane domains. Interestingly, this result also denotes differences between the active conformations produced by CaM and PA. To verify this point, we measured resonance energy transfer between PMCA labeled with eosin isothiocyanate at the ATP-binding site and the phospholipid RhoPE included in PMCA micelles. CaM decreased the efficiency of the energy transfer between these two probes, whereas PA did not. This result indicates that activation by CaM increases the distance between the ATP-binding site and the membrane, but PA does not affect this distance. Our results disclose main differences between PMCA conformations induced by CaM or PA and show that those differences involve transmembrane regions.
publishDate 2011
dc.date.none.fl_str_mv 2011-05
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/11336/18237
Mangialavori, Irene Cecilia; Villamil Giraldo, Ana María; Pignataro, María Florencia; Ferreira Gomes, Mariela Soledad; Caride, Ariel J.; et al.; Plasma Membrane Calcium Pump (PMCA) Differential Exposure of Hydrophobic Domains after Calmodulin and Phosphatidic Acid Activation; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry; 286; 21; 5-2011; 18397-18404
0021-9258
1083-351X
CONICET Digital
CONICET
url http://hdl.handle.net/11336/18237
identifier_str_mv Mangialavori, Irene Cecilia; Villamil Giraldo, Ana María; Pignataro, María Florencia; Ferreira Gomes, Mariela Soledad; Caride, Ariel J.; et al.; Plasma Membrane Calcium Pump (PMCA) Differential Exposure of Hydrophobic Domains after Calmodulin and Phosphatidic Acid Activation; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry; 286; 21; 5-2011; 18397-18404
0021-9258
1083-351X
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/pii/S0021925820510971
info:eu-repo/semantics/altIdentifier/doi/10.1074/jbc.M110.210088
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 Society for Biochemistry and Molecular Biology
publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
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 13.13397

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