P2X7-mediated ATP secretion is accompanied by depletion of cytosolic ATP
ATP and its metabolites are important extracellular signal transmitters acting on purinergic P2 and P1 receptors. Most cells can actively secrete ATP in response to a variety of external stimuli such as gating of the P2X7 receptor. We used Yac-1 murine lymphoma cells to study P2X7-mediated ATP relea...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Netherlands
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6635544/ https://www.ncbi.nlm.nih.gov/pubmed/31016551 http://dx.doi.org/10.1007/s11302-019-09654-5 |
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author | Johnsen, Bjarne Kaschubowski, Klaus Eric Nader, Sorush Schneider, Enja Nicola, Jan-Andrei Fliegert, Ralf Wolf, Insa M. A. Guse, Andreas H. Nikolaev, Viacheslav O. Koch-Nolte, Friedrich Haag, Friedrich |
author_facet | Johnsen, Bjarne Kaschubowski, Klaus Eric Nader, Sorush Schneider, Enja Nicola, Jan-Andrei Fliegert, Ralf Wolf, Insa M. A. Guse, Andreas H. Nikolaev, Viacheslav O. Koch-Nolte, Friedrich Haag, Friedrich |
author_sort | Johnsen, Bjarne |
collection | PubMed |
description | ATP and its metabolites are important extracellular signal transmitters acting on purinergic P2 and P1 receptors. Most cells can actively secrete ATP in response to a variety of external stimuli such as gating of the P2X7 receptor. We used Yac-1 murine lymphoma cells to study P2X7-mediated ATP release. These cells co-express P2X7 and ADP-ribosyltransferase ARTC2, permitting gating of P2X7 by NAD(+)-dependent ADP-ribosylation without the need to add exogenous ATP. Yac-1 cells released ATP into the extracellular space within minutes after stimulation with NAD(+). This was blocked by pre-incubation with the inhibitory P2X7-specific nanobody 13A7. Gating of P2X7 for 3 h significantly decreased intracellular ATP levels in living cells, but these had returned to normal by 20 h. P2X7-mediated ATP release was dependent on a rise in cytosolic calcium and the depletion of intracellular potassium, but was not blocked by inhibitors of pannexins or connexins. We used genetically encoded FRET-based ATP sensors targeted to the cytosol to image P2X7-mediated changes in the distribution of ATP in 3T3 fibroblasts co-expressing P2X7 and ARTC2 and in Yac-1 cells. In response to NAD(+), we observed a marked depletion of ATP in the cytosol. This study demonstrates the potential of ATP sensors as tools to study regulated ATP release by other cell types under other conditions. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s11302-019-09654-5) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-6635544 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Springer Netherlands |
record_format | MEDLINE/PubMed |
spelling | pubmed-66355442019-07-31 P2X7-mediated ATP secretion is accompanied by depletion of cytosolic ATP Johnsen, Bjarne Kaschubowski, Klaus Eric Nader, Sorush Schneider, Enja Nicola, Jan-Andrei Fliegert, Ralf Wolf, Insa M. A. Guse, Andreas H. Nikolaev, Viacheslav O. Koch-Nolte, Friedrich Haag, Friedrich Purinergic Signal Original Article ATP and its metabolites are important extracellular signal transmitters acting on purinergic P2 and P1 receptors. Most cells can actively secrete ATP in response to a variety of external stimuli such as gating of the P2X7 receptor. We used Yac-1 murine lymphoma cells to study P2X7-mediated ATP release. These cells co-express P2X7 and ADP-ribosyltransferase ARTC2, permitting gating of P2X7 by NAD(+)-dependent ADP-ribosylation without the need to add exogenous ATP. Yac-1 cells released ATP into the extracellular space within minutes after stimulation with NAD(+). This was blocked by pre-incubation with the inhibitory P2X7-specific nanobody 13A7. Gating of P2X7 for 3 h significantly decreased intracellular ATP levels in living cells, but these had returned to normal by 20 h. P2X7-mediated ATP release was dependent on a rise in cytosolic calcium and the depletion of intracellular potassium, but was not blocked by inhibitors of pannexins or connexins. We used genetically encoded FRET-based ATP sensors targeted to the cytosol to image P2X7-mediated changes in the distribution of ATP in 3T3 fibroblasts co-expressing P2X7 and ARTC2 and in Yac-1 cells. In response to NAD(+), we observed a marked depletion of ATP in the cytosol. This study demonstrates the potential of ATP sensors as tools to study regulated ATP release by other cell types under other conditions. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s11302-019-09654-5) contains supplementary material, which is available to authorized users. Springer Netherlands 2019-04-23 2019-06 /pmc/articles/PMC6635544/ /pubmed/31016551 http://dx.doi.org/10.1007/s11302-019-09654-5 Text en © The Author(s) 2019 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. |
spellingShingle | Original Article Johnsen, Bjarne Kaschubowski, Klaus Eric Nader, Sorush Schneider, Enja Nicola, Jan-Andrei Fliegert, Ralf Wolf, Insa M. A. Guse, Andreas H. Nikolaev, Viacheslav O. Koch-Nolte, Friedrich Haag, Friedrich P2X7-mediated ATP secretion is accompanied by depletion of cytosolic ATP |
title | P2X7-mediated ATP secretion is accompanied by depletion of cytosolic ATP |
title_full | P2X7-mediated ATP secretion is accompanied by depletion of cytosolic ATP |
title_fullStr | P2X7-mediated ATP secretion is accompanied by depletion of cytosolic ATP |
title_full_unstemmed | P2X7-mediated ATP secretion is accompanied by depletion of cytosolic ATP |
title_short | P2X7-mediated ATP secretion is accompanied by depletion of cytosolic ATP |
title_sort | p2x7-mediated atp secretion is accompanied by depletion of cytosolic atp |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6635544/ https://www.ncbi.nlm.nih.gov/pubmed/31016551 http://dx.doi.org/10.1007/s11302-019-09654-5 |
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