Kinetics of extracellular ATP in mastoparan 7-activated human erythrocytes
Background The peptide mastoparan 7 (MST7) stimulated ATP release in human erythrocytes. We explored intra- and 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 o...
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2013
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| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03044165_v1830_n10_p4692_LealDenis http://hdl.handle.net/20.500.12110/paper_03044165_v1830_n10_p4692_LealDenis |
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paper:paper_03044165_v1830_n10_p4692_LealDenis2023-06-08T15:29:52Z Kinetics of extracellular ATP in mastoparan 7-activated human erythrocytes ATP ATPases Erythrocyte Extracellular Pannexin 1 adenosine triphosphatase adenosine triphosphate carbenoxolone cell protein mastoparan mastoparan 7 pannexin 1 poly dextro lysine probenecid unclassified drug animal cell article cell activation cell swelling chemical reaction kinetics concentration (parameters) controlled study enzyme activity erythrocyte erythrocyte adhesiveness erythrocyte volume experimental dog hematocrit human human cell hydrolysis inhibition kinetics intracellular signaling long term exposure mouse nonhuman nucleotide transport priority journal reduction kinetics wild type 3V a cAMP activating cocktail containing isoproterenol, forskolin and papaverine ATPases ATPe ATPi carbenoxolone CBX cilostazol CTZ Erythrocyte extracellular ATP Extracellular ATP flux of ATP release flux of ATPe hydrolysis GEFs guanine exchange factors intracellular ATP J(R) J(V) mastoparan MST pannexin 1 Pannexin 1 pannexin 1 heterozygous mice pannexin 1 knockout mice, PTX, pertussis toxin pannexin 1 wild type mice PBC pnx pnx(+/+) pnx(+/-) pnx(-/-) probenecid rbcs red blood cells relative cell volume Vr Adenosine Triphosphate Animals Dogs Erythrocytes Humans Hydrolysis Kinetics Mice Mice, Knockout Peptides Signal Transduction Mus Background The peptide mastoparan 7 (MST7) stimulated ATP release in human erythrocytes. We explored intra- and 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. © 2013 Elsevier B.V. 2013 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03044165_v1830_n10_p4692_LealDenis http://hdl.handle.net/20.500.12110/paper_03044165_v1830_n10_p4692_LealDenis |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
ATP ATPases Erythrocyte Extracellular Pannexin 1 adenosine triphosphatase adenosine triphosphate carbenoxolone cell protein mastoparan mastoparan 7 pannexin 1 poly dextro lysine probenecid unclassified drug animal cell article cell activation cell swelling chemical reaction kinetics concentration (parameters) controlled study enzyme activity erythrocyte erythrocyte adhesiveness erythrocyte volume experimental dog hematocrit human human cell hydrolysis inhibition kinetics intracellular signaling long term exposure mouse nonhuman nucleotide transport priority journal reduction kinetics wild type 3V a cAMP activating cocktail containing isoproterenol, forskolin and papaverine ATPases ATPe ATPi carbenoxolone CBX cilostazol CTZ Erythrocyte extracellular ATP Extracellular ATP flux of ATP release flux of ATPe hydrolysis GEFs guanine exchange factors intracellular ATP J(R) J(V) mastoparan MST pannexin 1 Pannexin 1 pannexin 1 heterozygous mice pannexin 1 knockout mice, PTX, pertussis toxin pannexin 1 wild type mice PBC pnx pnx(+/+) pnx(+/-) pnx(-/-) probenecid rbcs red blood cells relative cell volume Vr Adenosine Triphosphate Animals Dogs Erythrocytes Humans Hydrolysis Kinetics Mice Mice, Knockout Peptides Signal Transduction Mus |
| spellingShingle |
ATP ATPases Erythrocyte Extracellular Pannexin 1 adenosine triphosphatase adenosine triphosphate carbenoxolone cell protein mastoparan mastoparan 7 pannexin 1 poly dextro lysine probenecid unclassified drug animal cell article cell activation cell swelling chemical reaction kinetics concentration (parameters) controlled study enzyme activity erythrocyte erythrocyte adhesiveness erythrocyte volume experimental dog hematocrit human human cell hydrolysis inhibition kinetics intracellular signaling long term exposure mouse nonhuman nucleotide transport priority journal reduction kinetics wild type 3V a cAMP activating cocktail containing isoproterenol, forskolin and papaverine ATPases ATPe ATPi carbenoxolone CBX cilostazol CTZ Erythrocyte extracellular ATP Extracellular ATP flux of ATP release flux of ATPe hydrolysis GEFs guanine exchange factors intracellular ATP J(R) J(V) mastoparan MST pannexin 1 Pannexin 1 pannexin 1 heterozygous mice pannexin 1 knockout mice, PTX, pertussis toxin pannexin 1 wild type mice PBC pnx pnx(+/+) pnx(+/-) pnx(-/-) probenecid rbcs red blood cells relative cell volume Vr Adenosine Triphosphate Animals Dogs Erythrocytes Humans Hydrolysis Kinetics Mice Mice, Knockout Peptides Signal Transduction Mus Kinetics of extracellular ATP in mastoparan 7-activated human erythrocytes |
| topic_facet |
ATP ATPases Erythrocyte Extracellular Pannexin 1 adenosine triphosphatase adenosine triphosphate carbenoxolone cell protein mastoparan mastoparan 7 pannexin 1 poly dextro lysine probenecid unclassified drug animal cell article cell activation cell swelling chemical reaction kinetics concentration (parameters) controlled study enzyme activity erythrocyte erythrocyte adhesiveness erythrocyte volume experimental dog hematocrit human human cell hydrolysis inhibition kinetics intracellular signaling long term exposure mouse nonhuman nucleotide transport priority journal reduction kinetics wild type 3V a cAMP activating cocktail containing isoproterenol, forskolin and papaverine ATPases ATPe ATPi carbenoxolone CBX cilostazol CTZ Erythrocyte extracellular ATP Extracellular ATP flux of ATP release flux of ATPe hydrolysis GEFs guanine exchange factors intracellular ATP J(R) J(V) mastoparan MST pannexin 1 Pannexin 1 pannexin 1 heterozygous mice pannexin 1 knockout mice, PTX, pertussis toxin pannexin 1 wild type mice PBC pnx pnx(+/+) pnx(+/-) pnx(-/-) probenecid rbcs red blood cells relative cell volume Vr Adenosine Triphosphate Animals Dogs Erythrocytes Humans Hydrolysis Kinetics Mice Mice, Knockout Peptides Signal Transduction Mus |
| description |
Background The peptide mastoparan 7 (MST7) stimulated ATP release in human erythrocytes. We explored intra- and 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. © 2013 Elsevier B.V. |
| title |
Kinetics of extracellular ATP in mastoparan 7-activated human erythrocytes |
| 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 |
| publishDate |
2013 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03044165_v1830_n10_p4692_LealDenis http://hdl.handle.net/20.500.12110/paper_03044165_v1830_n10_p4692_LealDenis |
| _version_ |
1768543846259490816 |