Kinetics of extracellular ATP in mastoparan 7-activated human erythrocytes

Autores
Leal Denis, Maria Florencia; Incicco, Juan Jeremías; Espelt, Maria Victoria; Verstraeten, Sandra Viviana; Pignataro, Omar Pedro; Lazarowski, Eduardo R.; Schwarzbaum, Pablo Julio
Año de publicación
2013
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Background: The peptide mastoparan 7 (MST7) stimulated ATP release in human erythrocytes. We explored intraand extracellular processes governing the time-dependent accumulation of extracellular ATP (i.e., ATPe kinetics). Methods: Human erythrocytes were treated with MST7 in the presence or absence of two blockers of pannexin 1. ATPe concentration was monitored by luciferin–luciferase based real-time luminometry. Results: Exposure of human erythrocytes to MST7 led to an acute increase in [ATPe], followed by a slower increase phase. ATPe kinetics reflected a strong activation of ATP efflux and a low rate of ATPe hydrolysis by ectoATPase activity. Enhancement of [ATPe] by MST7 required adhesion of erythrocytes to poly-D-lysin-coated coverslips, and correlated with a 31% increase of cAMP and 10% cell swelling. However, when MST7 was dissolved in a hyperosmotic medium to block cell swelling, ATPe accumulation was inhibited by 49%. Erythrocytes pre-exposure to 10 μM of either carbenoxolone or probenecid, two blockers of pannexin 1, exhibited a partial reduction of ATP efflux. Erythrocytes from pannexin 1 knockout mice exhibited similar ATPe kinetics as those of wild type mice erythrocytes exposed to pannexin 1 blockers. Conclusions: MST7 induced release of ATP required either cell adhesion or strong activation of cAMP synthesis. Part of this release required cell swelling. Kinetic analysis and a data driven model suggested that ATP efflux is mediated by two ATP conduits displaying different kinetics, with one conduit being fully blocked by pannexin 1 blockers. General significance: Kinetic analysis of extracellular ATP accumulation from human erythrocytes and potential effects on microcirculation.
Fil: Leal Denis, Maria Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Fisicoquímica Biológicas; Argentina
Fil: Incicco, Juan Jeremías. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Fisicoquímica Biológicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Biológica; Argentina
Fil: Espelt, Maria Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Fisicoquímica Biológicas; Argentina
Fil: Verstraeten, Sandra Viviana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Fisicoquímica Biológicas; Argentina
Fil: Pignataro, Omar Pedro. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina
Fil: Lazarowski, Eduardo R.. University of North Carolina at Chapel Hill. Cystic Fibrosis/Pulmonary Research and Treatment Center; Estados Unidos
Fil: Schwarzbaum, Pablo Julio. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Fisicoquímica Biológicas; Argentina
Materia
ATP
ATPASES
ERYTHROCYTE
EXTRACELLULAR
PANNEXIN 1
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:11336/2772
Acceder
id CONICETDig_01ea759d571afe3ed848ea96e1f002c2
oai_identifier_str oai:ri.conicet.gov.ar:11336/2772
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Kinetics of extracellular ATP in mastoparan 7-activated human erythrocytesLeal Denis, Maria FlorenciaIncicco, Juan JeremíasEspelt, Maria VictoriaVerstraeten, Sandra VivianaPignataro, Omar PedroLazarowski, Eduardo R.Schwarzbaum, Pablo JulioATPATPASESERYTHROCYTEEXTRACELLULARPANNEXIN 1https://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Background: The peptide mastoparan 7 (MST7) stimulated ATP release in human erythrocytes. We explored intraand extracellular processes governing the time-dependent accumulation of extracellular ATP (i.e., ATPe kinetics). Methods: Human erythrocytes were treated with MST7 in the presence or absence of two blockers of pannexin 1. ATPe concentration was monitored by luciferin–luciferase based real-time luminometry. Results: Exposure of human erythrocytes to MST7 led to an acute increase in [ATPe], followed by a slower increase phase. ATPe kinetics reflected a strong activation of ATP efflux and a low rate of ATPe hydrolysis by ectoATPase activity. Enhancement of [ATPe] by MST7 required adhesion of erythrocytes to poly-D-lysin-coated coverslips, and correlated with a 31% increase of cAMP and 10% cell swelling. However, when MST7 was dissolved in a hyperosmotic medium to block cell swelling, ATPe accumulation was inhibited by 49%. Erythrocytes pre-exposure to 10 μM of either carbenoxolone or probenecid, two blockers of pannexin 1, exhibited a partial reduction of ATP efflux. Erythrocytes from pannexin 1 knockout mice exhibited similar ATPe kinetics as those of wild type mice erythrocytes exposed to pannexin 1 blockers. Conclusions: MST7 induced release of ATP required either cell adhesion or strong activation of cAMP synthesis. Part of this release required cell swelling. Kinetic analysis and a data driven model suggested that ATP efflux is mediated by two ATP conduits displaying different kinetics, with one conduit being fully blocked by pannexin 1 blockers. General significance: Kinetic analysis of extracellular ATP accumulation from human erythrocytes and potential effects on microcirculation.Fil: Leal Denis, Maria Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Fisicoquímica Biológicas; ArgentinaFil: Incicco, Juan Jeremías. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Fisicoquímica Biológicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Biológica; ArgentinaFil: Espelt, Maria Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Fisicoquímica Biológicas; ArgentinaFil: Verstraeten, Sandra Viviana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Fisicoquímica Biológicas; ArgentinaFil: Pignataro, Omar Pedro. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); ArgentinaFil: Lazarowski, Eduardo R.. University of North Carolina at Chapel Hill. Cystic Fibrosis/Pulmonary Research and Treatment Center; Estados UnidosFil: Schwarzbaum, Pablo Julio. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Fisicoquímica Biológicas; ArgentinaElsevier Science2013-10info: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/11336/2772Leal Denis, Maria Florencia; Incicco, Juan Jeremías; Espelt, Maria Victoria; Verstraeten, Sandra Viviana; Pignataro, Omar Pedro; et al.; Kinetics of extracellular ATP in mastoparan 7-activated human erythrocytes; Elsevier Science; Biochimica et Biophysica Acta- General Subjects; 1830; 10; 10-2013; 4692-47070304-4165enginfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.bbagen.2013.05.033info:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S0304416513002298info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:33:43Zoai:ri.conicet.gov.ar:11336/2772instacron: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 09:33:44.196CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Kinetics of extracellular ATP in mastoparan 7-activated human erythrocytes
title Kinetics of extracellular ATP in mastoparan 7-activated human erythrocytes
spellingShingle Kinetics of extracellular ATP in mastoparan 7-activated human erythrocytes
Leal Denis, Maria Florencia
ATP
ATPASES
ERYTHROCYTE
EXTRACELLULAR
PANNEXIN 1
title_short Kinetics of extracellular ATP in mastoparan 7-activated human erythrocytes
title_full Kinetics of extracellular ATP in mastoparan 7-activated human erythrocytes
title_fullStr Kinetics of extracellular ATP in mastoparan 7-activated human erythrocytes
title_full_unstemmed Kinetics of extracellular ATP in mastoparan 7-activated human erythrocytes
title_sort Kinetics of extracellular ATP in mastoparan 7-activated human erythrocytes
dc.creator.none.fl_str_mv Leal Denis, Maria Florencia
Incicco, Juan Jeremías
Espelt, Maria Victoria
Verstraeten, Sandra Viviana
Pignataro, Omar Pedro
Lazarowski, Eduardo R.
Schwarzbaum, Pablo Julio
author Leal Denis, Maria Florencia
author_facet Leal Denis, Maria Florencia
Incicco, Juan Jeremías
Espelt, Maria Victoria
Verstraeten, Sandra Viviana
Pignataro, Omar Pedro
Lazarowski, Eduardo R.
Schwarzbaum, Pablo Julio
author_role author
author2 Incicco, Juan Jeremías
Espelt, Maria Victoria
Verstraeten, Sandra Viviana
Pignataro, Omar Pedro
Lazarowski, Eduardo R.
Schwarzbaum, Pablo Julio
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv ATP
ATPASES
ERYTHROCYTE
EXTRACELLULAR
PANNEXIN 1
topic ATP
ATPASES
ERYTHROCYTE
EXTRACELLULAR
PANNEXIN 1
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Background: The peptide mastoparan 7 (MST7) stimulated ATP release in human erythrocytes. We explored intraand extracellular processes governing the time-dependent accumulation of extracellular ATP (i.e., ATPe kinetics). Methods: Human erythrocytes were treated with MST7 in the presence or absence of two blockers of pannexin 1. ATPe concentration was monitored by luciferin–luciferase based real-time luminometry. Results: Exposure of human erythrocytes to MST7 led to an acute increase in [ATPe], followed by a slower increase phase. ATPe kinetics reflected a strong activation of ATP efflux and a low rate of ATPe hydrolysis by ectoATPase activity. Enhancement of [ATPe] by MST7 required adhesion of erythrocytes to poly-D-lysin-coated coverslips, and correlated with a 31% increase of cAMP and 10% cell swelling. However, when MST7 was dissolved in a hyperosmotic medium to block cell swelling, ATPe accumulation was inhibited by 49%. Erythrocytes pre-exposure to 10 μM of either carbenoxolone or probenecid, two blockers of pannexin 1, exhibited a partial reduction of ATP efflux. Erythrocytes from pannexin 1 knockout mice exhibited similar ATPe kinetics as those of wild type mice erythrocytes exposed to pannexin 1 blockers. Conclusions: MST7 induced release of ATP required either cell adhesion or strong activation of cAMP synthesis. Part of this release required cell swelling. Kinetic analysis and a data driven model suggested that ATP efflux is mediated by two ATP conduits displaying different kinetics, with one conduit being fully blocked by pannexin 1 blockers. General significance: Kinetic analysis of extracellular ATP accumulation from human erythrocytes and potential effects on microcirculation.
Fil: Leal Denis, Maria Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Fisicoquímica Biológicas; Argentina
Fil: Incicco, Juan Jeremías. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Fisicoquímica Biológicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Biológica; Argentina
Fil: Espelt, Maria Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Fisicoquímica Biológicas; Argentina
Fil: Verstraeten, Sandra Viviana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Fisicoquímica Biológicas; Argentina
Fil: Pignataro, Omar Pedro. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina
Fil: Lazarowski, Eduardo R.. University of North Carolina at Chapel Hill. Cystic Fibrosis/Pulmonary Research and Treatment Center; Estados Unidos
Fil: Schwarzbaum, Pablo Julio. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Fisicoquímica Biológicas; Argentina
description Background: The peptide mastoparan 7 (MST7) stimulated ATP release in human erythrocytes. We explored intraand extracellular processes governing the time-dependent accumulation of extracellular ATP (i.e., ATPe kinetics). Methods: Human erythrocytes were treated with MST7 in the presence or absence of two blockers of pannexin 1. ATPe concentration was monitored by luciferin–luciferase based real-time luminometry. Results: Exposure of human erythrocytes to MST7 led to an acute increase in [ATPe], followed by a slower increase phase. ATPe kinetics reflected a strong activation of ATP efflux and a low rate of ATPe hydrolysis by ectoATPase activity. Enhancement of [ATPe] by MST7 required adhesion of erythrocytes to poly-D-lysin-coated coverslips, and correlated with a 31% increase of cAMP and 10% cell swelling. However, when MST7 was dissolved in a hyperosmotic medium to block cell swelling, ATPe accumulation was inhibited by 49%. Erythrocytes pre-exposure to 10 μM of either carbenoxolone or probenecid, two blockers of pannexin 1, exhibited a partial reduction of ATP efflux. Erythrocytes from pannexin 1 knockout mice exhibited similar ATPe kinetics as those of wild type mice erythrocytes exposed to pannexin 1 blockers. Conclusions: MST7 induced release of ATP required either cell adhesion or strong activation of cAMP synthesis. Part of this release required cell swelling. Kinetic analysis and a data driven model suggested that ATP efflux is mediated by two ATP conduits displaying different kinetics, with one conduit being fully blocked by pannexin 1 blockers. General significance: Kinetic analysis of extracellular ATP accumulation from human erythrocytes and potential effects on microcirculation.
publishDate 2013
dc.date.none.fl_str_mv 2013-10
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/11336/2772
Leal Denis, Maria Florencia; Incicco, Juan Jeremías; Espelt, Maria Victoria; Verstraeten, Sandra Viviana; Pignataro, Omar Pedro; et al.; Kinetics of extracellular ATP in mastoparan 7-activated human erythrocytes; Elsevier Science; Biochimica et Biophysica Acta- General Subjects; 1830; 10; 10-2013; 4692-4707
0304-4165
url http://hdl.handle.net/11336/2772
identifier_str_mv Leal Denis, Maria Florencia; Incicco, Juan Jeremías; Espelt, Maria Victoria; Verstraeten, Sandra Viviana; Pignataro, Omar Pedro; et al.; Kinetics of extracellular ATP in mastoparan 7-activated human erythrocytes; Elsevier Science; Biochimica et Biophysica Acta- General Subjects; 1830; 10; 10-2013; 4692-4707
0304-4165
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1016/j.bbagen.2013.05.033
info:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S0304416513002298
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
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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