Monitoring cytosolic H2O2 fluctuations arising from altered plasma membrane gradients or from mitochondrial activity
| Autor: | José Ayté, Iria Medraño-Fernandez, Marta Moral-Blanch, Eric Bautista, Laura de Cubas, Susanna Boronat, Mercè Carmona, Roberto Sitia, Elena Hidalgo |
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| Přispěvatelé: | Carmona, M., de Cubas, L., Bautista, E., Moral-Blanch, M., Medrano-Fernandez, I., Sitia, R., Boronat, S., Ayte, J., Hidalgo, E. |
| Rok vydání: | 2019 |
| Předmět: |
inorganic chemicals
0301 basic medicine Science Molecular Probe Techniques General Physics and Astronomy Aquaporin Mitochondrion Protein Engineering General Biochemistry Genetics and Molecular Biology Stress signalling 03 medical and health sciences Cytosol Genes Reporter Extracellular lcsh:Science Multidisciplinary 030102 biochemistry & molecular biology Chemistry Cell Membrane Luminescent Protein Peroxiredoxin Hydrogen Peroxide General Chemistry Electron transport chain Mitochondria Molecular Probe 030104 developmental biology Biophysics lcsh:Q Chemical tools Oxidation-Reduction Schizosaccharomyce Intracellular |
| Zdroj: | Nature Communications, Vol 10, Iss 1, Pp 1-13 (2019) Recercat. Dipósit de la Recerca de Catalunya instname |
| ISSN: | 2041-1723 |
| DOI: | 10.1038/s41467-019-12475-0 |
| Popis: | Genetically encoded probes monitoring H2O2 fluctuations in living organisms are key to decipher redox signaling events. Here we use a new probe, roGFP2-Tpx1.C169S, to monitor pre-toxic fluctuations of peroxides in fission yeast, where the concentrations linked to signaling or to toxicity have been established. This probe is able to detect nanomolar fluctuations of intracellular H2O2 caused by extracellular peroxides; expression of human aquaporin 8 channels H2O2 entry into fission yeast decreasing membrane gradients. The probe also detects H2O2 bursts from mitochondria after addition of electron transport chain inhibitors, the extent of probe oxidation being proportional to the mitochondrial activity. The oxidation of this probe is an indicator of steady-state levels of H2O2 in different genetic backgrounds. Metabolic reprogramming during growth in low-glucose media causes probe reduction due to the activation of antioxidant cascades. We demonstrate how peroxiredoxin-based probes can be used to monitor physiological H2O2 fluctuations. This work is supported by the Ministerio de Ciencia, Innovación y Universidades (Spain), PLAN E and FEDER (BFU2015-68350-P and PGC2018-093920-B-I00 to E.H.) and by Unidad de Excelencia María de Maeztu (MDM-2014-0370). The Oxidative Stress and Cell Cycle group is also supported by Generalitat de Catalunya (Spain) (2017-SGR-539). E.H. is recipient of an ICREA Academia Award (Generalitat de Catalunya, Spain). |
| Databáze: | OpenAIRE |
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