Mechanosensitive Pannexin 1 Activity Is Modulated by Stomatin in Human Red Blood Cells
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 act...
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| Format: | Article |
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MDPI AG
2022-08-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/16/9401 |
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| author | Sarah Rougé Sandrine Genetet Maria Florencia Leal Denis Michael Dussiot Pablo Julio Schwarzbaum Mariano Anibal Ostuni Isabelle Mouro-Chanteloup |
| author_facet | Sarah Rougé Sandrine Genetet Maria Florencia Leal Denis Michael Dussiot Pablo Julio Schwarzbaum Mariano Anibal Ostuni Isabelle Mouro-Chanteloup |
| author_sort | Sarah Rougé |
| collection | DOAJ |
| 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<sup>+</sup> and stom<sup>−</sup> 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. |
| first_indexed | 2024-03-09T04:18:39Z |
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| id | doaj.art-17ed9e6f13094063b623ae8be3d506a3 |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T04:18:39Z |
| publishDate | 2022-08-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-17ed9e6f13094063b623ae8be3d506a32023-12-03T13:50:01ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-08-012316940110.3390/ijms23169401Mechanosensitive Pannexin 1 Activity Is Modulated by Stomatin in Human Red Blood CellsSarah Rougé0Sandrine Genetet1Maria Florencia Leal Denis2Michael Dussiot3Pablo Julio Schwarzbaum4Mariano Anibal Ostuni5Isabelle Mouro-Chanteloup6Université Paris Cité and Université des Antilles, INSERM U1134, BIGR, F-75014 Paris, FranceUniversité Paris Cité and Université des Antilles, INSERM U1134, BIGR, F-75014 Paris, FranceInstituto de Química y Fisico-Química Biológicas “Prof. Alejandro C. Paladini”, UBA, CONICET, Facultad de Farmacia y Bioquímica, 1113 Buenos Aires, ArgentinaUniversité Paris Cité, INSERM U1163, IMAGINE, F-75015 Paris, FranceInstituto de Química y Fisico-Química Biológicas “Prof. Alejandro C. Paladini”, UBA, CONICET, Facultad de Farmacia y Bioquímica, 1113 Buenos Aires, ArgentinaUniversité Paris Cité and Université des Antilles, INSERM U1134, BIGR, F-75014 Paris, FranceUniversité Paris Cité and Université des Antilles, INSERM U1134, BIGR, F-75014 Paris, FrancePannexin 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<sup>+</sup> and stom<sup>−</sup> 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.https://www.mdpi.com/1422-0067/23/16/9401erythrocyteATP releasepannexin activitystomatinoverhydrated hereditary stomatocytosis |
| spellingShingle | Sarah Rougé Sandrine Genetet Maria Florencia Leal Denis Michael Dussiot Pablo Julio Schwarzbaum Mariano Anibal Ostuni Isabelle Mouro-Chanteloup Mechanosensitive Pannexin 1 Activity Is Modulated by Stomatin in Human Red Blood Cells International Journal of Molecular Sciences erythrocyte ATP release pannexin activity stomatin overhydrated hereditary stomatocytosis |
| title | 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_short | 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 |
| topic | erythrocyte ATP release pannexin activity stomatin overhydrated hereditary stomatocytosis |
| url | https://www.mdpi.com/1422-0067/23/16/9401 |
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