Regulation of plasma membrane calcium ATPase (PMCA) by actin cytoskeleton
- Autores
- Vigil, Maximiliano Angel; Picco, María Elisa; Rinaldi, Debora Eugenia; Rossi, Rolando Carlos; Rey, Osvaldo; Rossi, Juan Pablo Francisco; Ferreira Gomes, Mariela Soledad
- Año de publicación
- 2021
- Idioma
- inglés
- Tipo de recurso
- documento de conferencia
- Estado
- versión publicada
- Descripción
- The Plasma Membrane Calcium ATPase (PMCA) is a calmodulin-modulated P-type ATPaseresponsible for the maintenance of low intracellular concentrations of Ca2+ in mosteukaryotic cells. Our group have previously shown that purified actin can exert a dualmodulation on the activity of Ca2+-ATPase 4b isoform (hPMCA4b): F-actin inhibits it whileshort actin oligomers may contribute to its activation. These studies had to be performedwith purified proteins given the nature of the biophysical and biochemical approachesused.On the other hand, in HEK293 human cells that overexpressed PMCA2w/b isoform, theactin depolymerization upon Cytochalasin D (CytD) treatment significantly increasedPMCA2-mediated Ca2+ extrusion and when F-actin was stabilized using jasplakinolide,PMCA2w/b activity was completely abolished.In order to assess whether the functional interaction between the hPMCA4 isoform and theactin cytoskeleton may be of physiological relevance, we decided to further characterize itin the context of a living cell by monitoring in real-time the changes in the actinpolymerization and cytosolic Ca2+ concentration ([Ca2+]cyt). For this, hPMCA4 isoform wastransiently expressed in HEK293T cells. The dynamics of [Ca2+]cyt was performed usingthe fluorescent probe Fluo-4 and studying the alterations in [Ca2+]cyt generated by Ca2+release from the endoplasmic reticulum, and by extracellular Ca2+ entry through store-operated Ca2+ channels. The dynamics of actin polymerization was performed transientlyexpressing LifeAct-Ruby.Results show that the alteration of actin polymerization by CytD treatment significantlyincreased hPMCA4 activity (102%). On the other hand, in absent of CytD, actinpolymerization dynamics did not change after TG stimulus, while after Ca2+ stimulus, anactin reorganization was observed. This reorganization takes place at the same times thatthe hPMCA4 increases its activity, suggesting that hPMCA4 may be activated by actindepolymerization in the cells.
Fil: Vigil, Maximiliano Angel. 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: Picco, María Elisa. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Inmunología, Genética y Metabolismo. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Inmunología, Genética y Metabolismo; Argentina
Fil: Rinaldi, Debora Eugenia. 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. 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: Rey, Osvaldo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Inmunología, Genética y Metabolismo. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Inmunología, Genética y Metabolismo; Argentina
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
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
XLIX Reunión Anual de la Sociedad Argentina de Biofísica
Argentina
Sociedad Argentina de Biofísica - Materia
-
PLASMA MEMBRANE CA2+-ATPASE
MODULATION
ACTIN - Nivel de accesibilidad
- acceso abierto
- Condiciones de uso
- https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
- Repositorio
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/173665
Ver los metadatos del registro completo
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Regulation of plasma membrane calcium ATPase (PMCA) by actin cytoskeletonVigil, Maximiliano AngelPicco, María ElisaRinaldi, Debora EugeniaRossi, Rolando CarlosRey, OsvaldoRossi, Juan Pablo FranciscoFerreira Gomes, Mariela SoledadPLASMA MEMBRANE CA2+-ATPASEMODULATIONACTINhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The Plasma Membrane Calcium ATPase (PMCA) is a calmodulin-modulated P-type ATPaseresponsible for the maintenance of low intracellular concentrations of Ca2+ in mosteukaryotic cells. Our group have previously shown that purified actin can exert a dualmodulation on the activity of Ca2+-ATPase 4b isoform (hPMCA4b): F-actin inhibits it whileshort actin oligomers may contribute to its activation. These studies had to be performedwith purified proteins given the nature of the biophysical and biochemical approachesused.On the other hand, in HEK293 human cells that overexpressed PMCA2w/b isoform, theactin depolymerization upon Cytochalasin D (CytD) treatment significantly increasedPMCA2-mediated Ca2+ extrusion and when F-actin was stabilized using jasplakinolide,PMCA2w/b activity was completely abolished.In order to assess whether the functional interaction between the hPMCA4 isoform and theactin cytoskeleton may be of physiological relevance, we decided to further characterize itin the context of a living cell by monitoring in real-time the changes in the actinpolymerization and cytosolic Ca2+ concentration ([Ca2+]cyt). For this, hPMCA4 isoform wastransiently expressed in HEK293T cells. The dynamics of [Ca2+]cyt was performed usingthe fluorescent probe Fluo-4 and studying the alterations in [Ca2+]cyt generated by Ca2+release from the endoplasmic reticulum, and by extracellular Ca2+ entry through store-operated Ca2+ channels. The dynamics of actin polymerization was performed transientlyexpressing LifeAct-Ruby.Results show that the alteration of actin polymerization by CytD treatment significantlyincreased hPMCA4 activity (102%). On the other hand, in absent of CytD, actinpolymerization dynamics did not change after TG stimulus, while after Ca2+ stimulus, anactin reorganization was observed. This reorganization takes place at the same times thatthe hPMCA4 increases its activity, suggesting that hPMCA4 may be activated by actindepolymerization in the cells.Fil: Vigil, Maximiliano Angel. 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: Picco, María Elisa. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Inmunología, Genética y Metabolismo. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Inmunología, Genética y Metabolismo; ArgentinaFil: Rinaldi, Debora Eugenia. 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. 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: Rey, Osvaldo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Inmunología, Genética y Metabolismo. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Inmunología, Genética y Metabolismo; ArgentinaFil: 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; 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; ArgentinaXLIX Reunión Anual de la Sociedad Argentina de BiofísicaArgentinaSociedad Argentina de BiofísicaSociedad Argentina de Biofísica2021info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectReuniónBookhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/173665Regulation of plasma membrane calcium ATPase (PMCA) by actin cytoskeleton; XLIX Reunión Anual de la Sociedad Argentina de Biofísica; Argentina; 2021; 50-50978-987-27591-9-3CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://biofisica.org.ar/reuniones-cientificas/Nacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:55:19Zoai:ri.conicet.gov.ar:11336/173665instacron: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 09:55:19.718CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse |
dc.title.none.fl_str_mv |
Regulation of plasma membrane calcium ATPase (PMCA) by actin cytoskeleton |
title |
Regulation of plasma membrane calcium ATPase (PMCA) by actin cytoskeleton |
spellingShingle |
Regulation of plasma membrane calcium ATPase (PMCA) by actin cytoskeleton Vigil, Maximiliano Angel PLASMA MEMBRANE CA2+-ATPASE MODULATION ACTIN |
title_short |
Regulation of plasma membrane calcium ATPase (PMCA) by actin cytoskeleton |
title_full |
Regulation of plasma membrane calcium ATPase (PMCA) by actin cytoskeleton |
title_fullStr |
Regulation of plasma membrane calcium ATPase (PMCA) by actin cytoskeleton |
title_full_unstemmed |
Regulation of plasma membrane calcium ATPase (PMCA) by actin cytoskeleton |
title_sort |
Regulation of plasma membrane calcium ATPase (PMCA) by actin cytoskeleton |
dc.creator.none.fl_str_mv |
Vigil, Maximiliano Angel Picco, María Elisa Rinaldi, Debora Eugenia Rossi, Rolando Carlos Rey, Osvaldo Rossi, Juan Pablo Francisco Ferreira Gomes, Mariela Soledad |
author |
Vigil, Maximiliano Angel |
author_facet |
Vigil, Maximiliano Angel Picco, María Elisa Rinaldi, Debora Eugenia Rossi, Rolando Carlos Rey, Osvaldo Rossi, Juan Pablo Francisco Ferreira Gomes, Mariela Soledad |
author_role |
author |
author2 |
Picco, María Elisa Rinaldi, Debora Eugenia Rossi, Rolando Carlos Rey, Osvaldo Rossi, Juan Pablo Francisco Ferreira Gomes, Mariela Soledad |
author2_role |
author author author author author author |
dc.subject.none.fl_str_mv |
PLASMA MEMBRANE CA2+-ATPASE MODULATION ACTIN |
topic |
PLASMA MEMBRANE CA2+-ATPASE MODULATION ACTIN |
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 Plasma Membrane Calcium ATPase (PMCA) is a calmodulin-modulated P-type ATPaseresponsible for the maintenance of low intracellular concentrations of Ca2+ in mosteukaryotic cells. Our group have previously shown that purified actin can exert a dualmodulation on the activity of Ca2+-ATPase 4b isoform (hPMCA4b): F-actin inhibits it whileshort actin oligomers may contribute to its activation. These studies had to be performedwith purified proteins given the nature of the biophysical and biochemical approachesused.On the other hand, in HEK293 human cells that overexpressed PMCA2w/b isoform, theactin depolymerization upon Cytochalasin D (CytD) treatment significantly increasedPMCA2-mediated Ca2+ extrusion and when F-actin was stabilized using jasplakinolide,PMCA2w/b activity was completely abolished.In order to assess whether the functional interaction between the hPMCA4 isoform and theactin cytoskeleton may be of physiological relevance, we decided to further characterize itin the context of a living cell by monitoring in real-time the changes in the actinpolymerization and cytosolic Ca2+ concentration ([Ca2+]cyt). For this, hPMCA4 isoform wastransiently expressed in HEK293T cells. The dynamics of [Ca2+]cyt was performed usingthe fluorescent probe Fluo-4 and studying the alterations in [Ca2+]cyt generated by Ca2+release from the endoplasmic reticulum, and by extracellular Ca2+ entry through store-operated Ca2+ channels. The dynamics of actin polymerization was performed transientlyexpressing LifeAct-Ruby.Results show that the alteration of actin polymerization by CytD treatment significantlyincreased hPMCA4 activity (102%). On the other hand, in absent of CytD, actinpolymerization dynamics did not change after TG stimulus, while after Ca2+ stimulus, anactin reorganization was observed. This reorganization takes place at the same times thatthe hPMCA4 increases its activity, suggesting that hPMCA4 may be activated by actindepolymerization in the cells. Fil: Vigil, Maximiliano Angel. 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: Picco, María Elisa. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Inmunología, Genética y Metabolismo. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Inmunología, Genética y Metabolismo; Argentina Fil: Rinaldi, Debora Eugenia. 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. 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: Rey, Osvaldo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Inmunología, Genética y Metabolismo. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Inmunología, Genética y Metabolismo; Argentina 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 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 XLIX Reunión Anual de la Sociedad Argentina de Biofísica Argentina Sociedad Argentina de Biofísica |
description |
The Plasma Membrane Calcium ATPase (PMCA) is a calmodulin-modulated P-type ATPaseresponsible for the maintenance of low intracellular concentrations of Ca2+ in mosteukaryotic cells. Our group have previously shown that purified actin can exert a dualmodulation on the activity of Ca2+-ATPase 4b isoform (hPMCA4b): F-actin inhibits it whileshort actin oligomers may contribute to its activation. These studies had to be performedwith purified proteins given the nature of the biophysical and biochemical approachesused.On the other hand, in HEK293 human cells that overexpressed PMCA2w/b isoform, theactin depolymerization upon Cytochalasin D (CytD) treatment significantly increasedPMCA2-mediated Ca2+ extrusion and when F-actin was stabilized using jasplakinolide,PMCA2w/b activity was completely abolished.In order to assess whether the functional interaction between the hPMCA4 isoform and theactin cytoskeleton may be of physiological relevance, we decided to further characterize itin the context of a living cell by monitoring in real-time the changes in the actinpolymerization and cytosolic Ca2+ concentration ([Ca2+]cyt). For this, hPMCA4 isoform wastransiently expressed in HEK293T cells. The dynamics of [Ca2+]cyt was performed usingthe fluorescent probe Fluo-4 and studying the alterations in [Ca2+]cyt generated by Ca2+release from the endoplasmic reticulum, and by extracellular Ca2+ entry through store-operated Ca2+ channels. The dynamics of actin polymerization was performed transientlyexpressing LifeAct-Ruby.Results show that the alteration of actin polymerization by CytD treatment significantlyincreased hPMCA4 activity (102%). On the other hand, in absent of CytD, actinpolymerization dynamics did not change after TG stimulus, while after Ca2+ stimulus, anactin reorganization was observed. This reorganization takes place at the same times thatthe hPMCA4 increases its activity, suggesting that hPMCA4 may be activated by actindepolymerization in the cells. |
publishDate |
2021 |
dc.date.none.fl_str_mv |
2021 |
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info:eu-repo/semantics/publishedVersion info:eu-repo/semantics/conferenceObject Reunión Book http://purl.org/coar/resource_type/c_5794 info:ar-repo/semantics/documentoDeConferencia |
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http://hdl.handle.net/11336/173665 Regulation of plasma membrane calcium ATPase (PMCA) by actin cytoskeleton; XLIX Reunión Anual de la Sociedad Argentina de Biofísica; Argentina; 2021; 50-50 978-987-27591-9-3 CONICET Digital CONICET |
url |
http://hdl.handle.net/11336/173665 |
identifier_str_mv |
Regulation of plasma membrane calcium ATPase (PMCA) by actin cytoskeleton; XLIX Reunión Anual de la Sociedad Argentina de Biofísica; Argentina; 2021; 50-50 978-987-27591-9-3 CONICET Digital CONICET |
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eng |
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eng |
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Nacional |
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Sociedad Argentina de Biofísica |
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Sociedad Argentina de Biofísica |
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dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar |
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