Extracellular ATP hydrolysis in Caco-2 human intestinal cell line
Extracellular nucleotides and nucleosides activate signaling pathways that play major roles in the physiology and pathophysiology of the gastrointestinal tract. Ectonucleotidases hydrolyze extracellular nucleotides and thus regulate ligand exposure to purinergic receptors. In this study, we investig...
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| Formato: | Articulo |
| Lenguaje: | Inglés |
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2021
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| Acceso en línea: | http://sedici.unlp.edu.ar/handle/10915/162850 |
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I19-R120-10915-1628502024-02-19T20:09:03Z http://sedici.unlp.edu.ar/handle/10915/162850 Extracellular ATP hydrolysis in Caco-2 human intestinal cell line Schachter, Julieta Alvarez, C. L. Bazzi, Z. Faillace, M. P. Corradi, G. Hattab, C. Rinaldi, D. E. Gonzalez Lebrero, R. Pucci Molineris, Melisa Eliana Sévigny, J. Ostuni, M. A. Schwarzbaum, P. J. 2021-10-01 2024-02-19T16:22:06Z en Ciencias Médicas ATP release ectoATPase Ectonucleotidases Extracellular ATP Intestinal epithelial cell Purinergic signaling Extracellular nucleotides and nucleosides activate signaling pathways that play major roles in the physiology and pathophysiology of the gastrointestinal tract. Ectonucleotidases hydrolyze extracellular nucleotides and thus regulate ligand exposure to purinergic receptors. In this study, we investigated the expression, localization and activities of ectonucleotidases using Caco-2 cells, a model of human intestinal epithelial cells. In addition, by studying ATP release and the rates of extracellular ATP (eATP) hydrolysis, we analyzed the contribution of these processes to the regulation of eATP in these cells. Results show that Caco-2 cells regulate the metabolism of eATP and by-products by ecto-nucleoside triphosphate diphosphohydrolase-1 and -2, a neutral ecto-phosphatase and ecto-5′-nucleotidase. All these ectoenzymes were kinetically characterized using intact cells, and their presence confirmed by denatured and native gels, western blot and cytoimmunofluorescence techniques. In addition, regulation of eATP was studied by monitoring the dynamic balance between intracellular ATP release and ectoATPase activity. Following mechanical and hypotonic stimuli, Caco-2 cells triggered a strong but transient release of intracellular ATP, with almost no energy cost, leading to a steep increase of eATP concentration, which was later reduced by ectoATPase activity. A data-driven algorithm allowed quantifying and predicting the rates of ATP release and ATP consumption contributing to the dynamic accumulation of ATP at the cell surface. Consejo Nacional de Investigaciones Científicas y Técnicas Instituto de Investigaciones Bioquímicas de La Plata Articulo Articulo http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf |
| institution |
Universidad Nacional de La Plata |
| institution_str |
I-19 |
| repository_str |
R-120 |
| collection |
SEDICI (UNLP) |
| language |
Inglés |
| topic |
Ciencias Médicas ATP release ectoATPase Ectonucleotidases Extracellular ATP Intestinal epithelial cell Purinergic signaling |
| spellingShingle |
Ciencias Médicas ATP release ectoATPase Ectonucleotidases Extracellular ATP Intestinal epithelial cell Purinergic signaling Schachter, Julieta Alvarez, C. L. Bazzi, Z. Faillace, M. P. Corradi, G. Hattab, C. Rinaldi, D. E. Gonzalez Lebrero, R. Pucci Molineris, Melisa Eliana Sévigny, J. Ostuni, M. A. Schwarzbaum, P. J. Extracellular ATP hydrolysis in Caco-2 human intestinal cell line |
| topic_facet |
Ciencias Médicas ATP release ectoATPase Ectonucleotidases Extracellular ATP Intestinal epithelial cell Purinergic signaling |
| description |
Extracellular nucleotides and nucleosides activate signaling pathways that play major roles in the physiology and pathophysiology of the gastrointestinal tract. Ectonucleotidases hydrolyze extracellular nucleotides and thus regulate ligand exposure to purinergic receptors. In this study, we investigated the expression, localization and activities of ectonucleotidases using Caco-2 cells, a model of human intestinal epithelial cells. In addition, by studying ATP release and the rates of extracellular ATP (eATP) hydrolysis, we analyzed the contribution of these processes to the regulation of eATP in these cells. Results show that Caco-2 cells regulate the metabolism of eATP and by-products by ecto-nucleoside triphosphate diphosphohydrolase-1 and -2, a neutral ecto-phosphatase and ecto-5′-nucleotidase. All these ectoenzymes were kinetically characterized using intact cells, and their presence confirmed by denatured and native gels, western blot and cytoimmunofluorescence techniques.
In addition, regulation of eATP was studied by monitoring the dynamic balance between intracellular ATP release and ectoATPase activity. Following mechanical and hypotonic stimuli, Caco-2 cells triggered a strong but transient release of intracellular ATP, with almost no energy cost, leading to a steep increase of eATP concentration, which was later reduced by ectoATPase activity. A data-driven algorithm allowed quantifying and predicting the rates of ATP release and ATP consumption contributing to the dynamic accumulation of ATP at the cell surface. |
| format |
Articulo Articulo |
| author |
Schachter, Julieta Alvarez, C. L. Bazzi, Z. Faillace, M. P. Corradi, G. Hattab, C. Rinaldi, D. E. Gonzalez Lebrero, R. Pucci Molineris, Melisa Eliana Sévigny, J. Ostuni, M. A. Schwarzbaum, P. J. |
| author_facet |
Schachter, Julieta Alvarez, C. L. Bazzi, Z. Faillace, M. P. Corradi, G. Hattab, C. Rinaldi, D. E. Gonzalez Lebrero, R. Pucci Molineris, Melisa Eliana Sévigny, J. Ostuni, M. A. Schwarzbaum, P. J. |
| author_sort |
Schachter, Julieta |
| title |
Extracellular ATP hydrolysis in Caco-2 human intestinal cell line |
| title_short |
Extracellular ATP hydrolysis in Caco-2 human intestinal cell line |
| title_full |
Extracellular ATP hydrolysis in Caco-2 human intestinal cell line |
| title_fullStr |
Extracellular ATP hydrolysis in Caco-2 human intestinal cell line |
| title_full_unstemmed |
Extracellular ATP hydrolysis in Caco-2 human intestinal cell line |
| title_sort |
extracellular atp hydrolysis in caco-2 human intestinal cell line |
| publishDate |
2021 |
| url |
http://sedici.unlp.edu.ar/handle/10915/162850 |
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| _version_ |
1807222465002209280 |