Mechanosensitive Pannexin 1 Activity Is Modulated by Stomatin in Human Red Blood Cells

Autores
Rougé, Sarah; Genetet, Sandrine; Leal Denis, Maria Florencia; Dussiot, Michael; Schwarzbaum, Pablo Julio; Ostuni, Mariano; Mouro Chanteloup, Isabelle
Año de publicación
2022
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Pannexin 1 (PANX1) was proposed to drive ATP release from red blood cells (RBCs) in response to stress conditions. Stomatin, a membrane protein regulating mechanosensitive channels, has been proposed to modulate PANX1 activity in non-erythroid cells. To determine whether stomatin modulates PANX1 activity in an erythroid context, we have (i) assessed the in situ stomatin-PANX1 interaction in RBCs, (ii) measured PANX1-stimulated activity in RBCs expressing stomatin or from OverHydrated Hereditary Stomatocytosis (OHSt) patients lacking stomatin, and in erythroid K562 cells invalidated for stomatin. Proximity Ligation Assay coupled with flow imaging shows 27.09% and 6.13% positive events in control and OHSt RBCs, respectively. The uptake of dyes 5(6)-Carboxyfluorescein (CF) and TO-PRO-3 was used to evaluate PANX1 activity. RBC permeability for CF is 34% and 11.8% in control and OHSt RBCs, respectively. PANX1 permeability for TO-PRO-3 is 35.72% and 18.42% in K562 stom+ and stom− clones, respectively. These results suggest an interaction between PANX1 and stomatin in human RBCs and show a significant defect in PANX1 activity in the absence of stomatin. Based on these results, we propose that stomatin plays a major role in opening the PANX1 pore by being involved in a caspase-independent lifting of autoinhibition.
Fil: Rougé, Sarah. Universite de Paris; Francia
Fil: Genetet, Sandrine. Universite de Paris; Francia
Fil: Leal Denis, Maria 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: Dussiot, Michael. Universite de Paris; Francia
Fil: Schwarzbaum, Pablo Julio. 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: Ostuni, Mariano. Universite de Paris; Francia
Fil: Mouro Chanteloup, Isabelle. Universite de Paris; Francia
Materia
ATP RELEASE
ERYTHROCYTE
OVERHYDRATED HEREDITARY STOMATOCYTOSIS
PANNEXIN ACTIVITY
STOMATIN
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/213630
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/213630
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Mechanosensitive Pannexin 1 Activity Is Modulated by Stomatin in Human Red Blood CellsRougé, SarahGenetet, SandrineLeal Denis, Maria FlorenciaDussiot, MichaelSchwarzbaum, Pablo JulioOstuni, MarianoMouro Chanteloup, IsabelleATP RELEASEERYTHROCYTEOVERHYDRATED HEREDITARY STOMATOCYTOSISPANNEXIN ACTIVITYSTOMATINhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Pannexin 1 (PANX1) was proposed to drive ATP release from red blood cells (RBCs) in response to stress conditions. Stomatin, a membrane protein regulating mechanosensitive channels, has been proposed to modulate PANX1 activity in non-erythroid cells. To determine whether stomatin modulates PANX1 activity in an erythroid context, we have (i) assessed the in situ stomatin-PANX1 interaction in RBCs, (ii) measured PANX1-stimulated activity in RBCs expressing stomatin or from OverHydrated Hereditary Stomatocytosis (OHSt) patients lacking stomatin, and in erythroid K562 cells invalidated for stomatin. Proximity Ligation Assay coupled with flow imaging shows 27.09% and 6.13% positive events in control and OHSt RBCs, respectively. The uptake of dyes 5(6)-Carboxyfluorescein (CF) and TO-PRO-3 was used to evaluate PANX1 activity. RBC permeability for CF is 34% and 11.8% in control and OHSt RBCs, respectively. PANX1 permeability for TO-PRO-3 is 35.72% and 18.42% in K562 stom+ and stom− clones, respectively. These results suggest an interaction between PANX1 and stomatin in human RBCs and show a significant defect in PANX1 activity in the absence of stomatin. Based on these results, we propose that stomatin plays a major role in opening the PANX1 pore by being involved in a caspase-independent lifting of autoinhibition.Fil: Rougé, Sarah. Universite de Paris; FranciaFil: Genetet, Sandrine. Universite de Paris; FranciaFil: Leal Denis, Maria 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: Dussiot, Michael. Universite de Paris; FranciaFil: Schwarzbaum, Pablo Julio. 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: Ostuni, Mariano. Universite de Paris; FranciaFil: Mouro Chanteloup, Isabelle. Universite de Paris; FranciaMolecular Diversity Preservation International2022-08info: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/11336/213630Rougé, Sarah; Genetet, Sandrine; Leal Denis, Maria Florencia; Dussiot, Michael; Schwarzbaum, Pablo Julio; et al.; Mechanosensitive Pannexin 1 Activity Is Modulated by Stomatin in Human Red Blood Cells; Molecular Diversity Preservation International; International Journal of Molecular Sciences; 23; 16; 8-2022; 1-191422-0067CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.mdpi.com/1422-0067/23/16/9401info:eu-repo/semantics/altIdentifier/doi/10.3390/ijms23169401info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:30:27Zoai:ri.conicet.gov.ar:11336/213630instacron: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 10:30:27.903CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Mechanosensitive Pannexin 1 Activity Is Modulated by Stomatin in Human Red Blood Cells
title Mechanosensitive Pannexin 1 Activity Is Modulated by Stomatin in Human Red Blood Cells
spellingShingle Mechanosensitive Pannexin 1 Activity Is Modulated by Stomatin in Human Red Blood Cells
Rougé, Sarah
ATP RELEASE
ERYTHROCYTE
OVERHYDRATED HEREDITARY STOMATOCYTOSIS
PANNEXIN ACTIVITY
STOMATIN
title_short Mechanosensitive Pannexin 1 Activity Is Modulated by Stomatin in Human Red Blood Cells
title_full Mechanosensitive Pannexin 1 Activity Is Modulated by Stomatin in Human Red Blood Cells
title_fullStr Mechanosensitive Pannexin 1 Activity Is Modulated by Stomatin in Human Red Blood Cells
title_full_unstemmed Mechanosensitive Pannexin 1 Activity Is Modulated by Stomatin in Human Red Blood Cells
title_sort Mechanosensitive Pannexin 1 Activity Is Modulated by Stomatin in Human Red Blood Cells
dc.creator.none.fl_str_mv Rougé, Sarah
Genetet, Sandrine
Leal Denis, Maria Florencia
Dussiot, Michael
Schwarzbaum, Pablo Julio
Ostuni, Mariano
Mouro Chanteloup, Isabelle
author Rougé, Sarah
author_facet Rougé, Sarah
Genetet, Sandrine
Leal Denis, Maria Florencia
Dussiot, Michael
Schwarzbaum, Pablo Julio
Ostuni, Mariano
Mouro Chanteloup, Isabelle
author_role author
author2 Genetet, Sandrine
Leal Denis, Maria Florencia
Dussiot, Michael
Schwarzbaum, Pablo Julio
Ostuni, Mariano
Mouro Chanteloup, Isabelle
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv ATP RELEASE
ERYTHROCYTE
OVERHYDRATED HEREDITARY STOMATOCYTOSIS
PANNEXIN ACTIVITY
STOMATIN
topic ATP RELEASE
ERYTHROCYTE
OVERHYDRATED HEREDITARY STOMATOCYTOSIS
PANNEXIN ACTIVITY
STOMATIN
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Pannexin 1 (PANX1) was proposed to drive ATP release from red blood cells (RBCs) in response to stress conditions. Stomatin, a membrane protein regulating mechanosensitive channels, has been proposed to modulate PANX1 activity in non-erythroid cells. To determine whether stomatin modulates PANX1 activity in an erythroid context, we have (i) assessed the in situ stomatin-PANX1 interaction in RBCs, (ii) measured PANX1-stimulated activity in RBCs expressing stomatin or from OverHydrated Hereditary Stomatocytosis (OHSt) patients lacking stomatin, and in erythroid K562 cells invalidated for stomatin. Proximity Ligation Assay coupled with flow imaging shows 27.09% and 6.13% positive events in control and OHSt RBCs, respectively. The uptake of dyes 5(6)-Carboxyfluorescein (CF) and TO-PRO-3 was used to evaluate PANX1 activity. RBC permeability for CF is 34% and 11.8% in control and OHSt RBCs, respectively. PANX1 permeability for TO-PRO-3 is 35.72% and 18.42% in K562 stom+ and stom− clones, respectively. These results suggest an interaction between PANX1 and stomatin in human RBCs and show a significant defect in PANX1 activity in the absence of stomatin. Based on these results, we propose that stomatin plays a major role in opening the PANX1 pore by being involved in a caspase-independent lifting of autoinhibition.
Fil: Rougé, Sarah. Universite de Paris; Francia
Fil: Genetet, Sandrine. Universite de Paris; Francia
Fil: Leal Denis, Maria 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: Dussiot, Michael. Universite de Paris; Francia
Fil: Schwarzbaum, Pablo Julio. 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: Ostuni, Mariano. Universite de Paris; Francia
Fil: Mouro Chanteloup, Isabelle. Universite de Paris; Francia
description Pannexin 1 (PANX1) was proposed to drive ATP release from red blood cells (RBCs) in response to stress conditions. Stomatin, a membrane protein regulating mechanosensitive channels, has been proposed to modulate PANX1 activity in non-erythroid cells. To determine whether stomatin modulates PANX1 activity in an erythroid context, we have (i) assessed the in situ stomatin-PANX1 interaction in RBCs, (ii) measured PANX1-stimulated activity in RBCs expressing stomatin or from OverHydrated Hereditary Stomatocytosis (OHSt) patients lacking stomatin, and in erythroid K562 cells invalidated for stomatin. Proximity Ligation Assay coupled with flow imaging shows 27.09% and 6.13% positive events in control and OHSt RBCs, respectively. The uptake of dyes 5(6)-Carboxyfluorescein (CF) and TO-PRO-3 was used to evaluate PANX1 activity. RBC permeability for CF is 34% and 11.8% in control and OHSt RBCs, respectively. PANX1 permeability for TO-PRO-3 is 35.72% and 18.42% in K562 stom+ and stom− clones, respectively. These results suggest an interaction between PANX1 and stomatin in human RBCs and show a significant defect in PANX1 activity in the absence of stomatin. Based on these results, we propose that stomatin plays a major role in opening the PANX1 pore by being involved in a caspase-independent lifting of autoinhibition.
publishDate 2022
dc.date.none.fl_str_mv 2022-08
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/11336/213630
Rougé, Sarah; Genetet, Sandrine; Leal Denis, Maria Florencia; Dussiot, Michael; Schwarzbaum, Pablo Julio; et al.; Mechanosensitive Pannexin 1 Activity Is Modulated by Stomatin in Human Red Blood Cells; Molecular Diversity Preservation International; International Journal of Molecular Sciences; 23; 16; 8-2022; 1-19
1422-0067
CONICET Digital
CONICET
url http://hdl.handle.net/11336/213630
identifier_str_mv Rougé, Sarah; Genetet, Sandrine; Leal Denis, Maria Florencia; Dussiot, Michael; Schwarzbaum, Pablo Julio; et al.; Mechanosensitive Pannexin 1 Activity Is Modulated by Stomatin in Human Red Blood Cells; Molecular Diversity Preservation International; International Journal of Molecular Sciences; 23; 16; 8-2022; 1-19
1422-0067
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.mdpi.com/1422-0067/23/16/9401
info:eu-repo/semantics/altIdentifier/doi/10.3390/ijms23169401
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Molecular Diversity Preservation International
publisher.none.fl_str_mv Molecular Diversity Preservation International
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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