The mitochondria-to-cytosol H2O2 gradient is caused by peroxiredoxin-dependent cytosolic scavenging

Fluorescent protein-based reporters used to measure intracellular H2O2 were developed to overcome the limitations of small permeable dyes. The two major families of genetically encoded redox reporters are the reduction-oxidation sensitive green fluorescent protein (roGFP)-based proteins fused to per...

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Autores: de Cubas, Laura, Pak, Valeriy V., Belousov, Vsevolod V., Ayté del Olmo, José, Hidalgo Hernando, Elena
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2021
País:España
Institución:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
Repositorio:Recercat. Dipósit de la Recerca de Catalunya
OAI Identifier:oai:recercat.cat:10230/47747
Acceso en línea:http://hdl.handle.net/10230/47747
http://dx.doi.org/10.3390/antiox10050731
Access Level:acceso abierto
Palabra clave:H2O2 gradients
HyPer7
Genetically encoded fluorescent H2O2 reporter
Mitochondria
roGFP-Tpx1.C169S
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spelling The mitochondria-to-cytosol H2O2 gradient is caused by peroxiredoxin-dependent cytosolic scavengingde Cubas, LauraPak, Valeriy V.Belousov, Vsevolod V.Ayté del Olmo, JoséHidalgo Hernando, ElenaH2O2 gradientsHyPer7Genetically encoded fluorescent H2O2 reporterMitochondriaroGFP-Tpx1.C169SFluorescent protein-based reporters used to measure intracellular H2O2 were developed to overcome the limitations of small permeable dyes. The two major families of genetically encoded redox reporters are the reduction-oxidation sensitive green fluorescent protein (roGFP)-based proteins fused to peroxiredoxins and HyPer and derivatives. We have used the most sensitive probes of each family, roGFP2-Tpx1.C169S and HyPer7, to monitor steady-state and fluctuating levels of peroxides in fission yeast. While both are able to monitor the nanomolar fluctuations of intracellular H2O2, the former is two-five times more sensitive than HyPer7, and roGFP2-Tpx1.C169S is partially oxidized in the cytosol of wild-type cells while HyPer7 is fully reduced. We have successfully expressed HyPer7 in the mitochondrial matrix, and it is ~40% oxidized, suggesting higher steady-state levels of peroxides, in the low micromolar range, than in the cytosol. Cytosolic HyPer7 can detect negligible H2O2 in the cytosol from mitochondrial origin unless the main H2O2 scavenger, the cytosolic peroxiredoxin Tpx1, is absent, while mitochondrial HyPer7 is oxidized to the same extent in wild-type and ∆tpx1 cells. We conclude that there is a bidirectional flux of H2O2 across the matrix and the cytosol, but Tpx1 rapidly and efficiently scavenges mitochondrial-generated peroxides and stops their steady-state cytosolic levels rising.This work is supported by the Ministerio de Ciencia, Innovación y Universidades (Spain),PLAN E and FEDER (PGC2018-093920-B-I00 to E.H., PGC2018-097248-B-I00 to J.A. and Redox Biologyand Medicine Research Network (Red2018-102576-T) to E.H.), and from the Ministry of Science andHigher Education (Russia) (075-15-2019-1933 to V.V.B.). The Oxidative Stress and Cell Cycle group isalso supported by Generalitat de Catalunya (Spain) (2017-SGR-539) and by Unidad de ExcelenciaMaría de Maeztu, funded by the AEI (CEX2018-000792-M) (Spain). L.d.C. is recipient of a María deMaeztu predoctoral fellowship from the Ministerio de Economía y Competitividad (Spain).MDPI202120212021info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfapplication/pdfhttp://hdl.handle.net/10230/47747http://dx.doi.org/10.3390/antiox10050731reponame:Recercat. Dipósit de la Recerca de Catalunyainstname:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)InglésAntioxidants (Basel). 2021;10(5):731info:eu-repo/grantAgreement/ES/2PE/PGC2018-093920-B-I00info:eu-repo/grantAgreement/ES/2PE/PGC2018-097248-B-I00© 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).https://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:recercat.cat:10230/477472026-05-29T05:05:01Z
dc.title.none.fl_str_mv The mitochondria-to-cytosol H2O2 gradient is caused by peroxiredoxin-dependent cytosolic scavenging
title The mitochondria-to-cytosol H2O2 gradient is caused by peroxiredoxin-dependent cytosolic scavenging
spellingShingle The mitochondria-to-cytosol H2O2 gradient is caused by peroxiredoxin-dependent cytosolic scavenging
de Cubas, Laura
H2O2 gradients
HyPer7
Genetically encoded fluorescent H2O2 reporter
Mitochondria
roGFP-Tpx1.C169S
title_short The mitochondria-to-cytosol H2O2 gradient is caused by peroxiredoxin-dependent cytosolic scavenging
title_full The mitochondria-to-cytosol H2O2 gradient is caused by peroxiredoxin-dependent cytosolic scavenging
title_fullStr The mitochondria-to-cytosol H2O2 gradient is caused by peroxiredoxin-dependent cytosolic scavenging
title_full_unstemmed The mitochondria-to-cytosol H2O2 gradient is caused by peroxiredoxin-dependent cytosolic scavenging
title_sort The mitochondria-to-cytosol H2O2 gradient is caused by peroxiredoxin-dependent cytosolic scavenging
dc.creator.none.fl_str_mv de Cubas, Laura
Pak, Valeriy V.
Belousov, Vsevolod V.
Ayté del Olmo, José
Hidalgo Hernando, Elena
author de Cubas, Laura
author_facet de Cubas, Laura
Pak, Valeriy V.
Belousov, Vsevolod V.
Ayté del Olmo, José
Hidalgo Hernando, Elena
author_role author
author2 Pak, Valeriy V.
Belousov, Vsevolod V.
Ayté del Olmo, José
Hidalgo Hernando, Elena
author2_role author
author
author
author
dc.subject.none.fl_str_mv H2O2 gradients
HyPer7
Genetically encoded fluorescent H2O2 reporter
Mitochondria
roGFP-Tpx1.C169S
topic H2O2 gradients
HyPer7
Genetically encoded fluorescent H2O2 reporter
Mitochondria
roGFP-Tpx1.C169S
description Fluorescent protein-based reporters used to measure intracellular H2O2 were developed to overcome the limitations of small permeable dyes. The two major families of genetically encoded redox reporters are the reduction-oxidation sensitive green fluorescent protein (roGFP)-based proteins fused to peroxiredoxins and HyPer and derivatives. We have used the most sensitive probes of each family, roGFP2-Tpx1.C169S and HyPer7, to monitor steady-state and fluctuating levels of peroxides in fission yeast. While both are able to monitor the nanomolar fluctuations of intracellular H2O2, the former is two-five times more sensitive than HyPer7, and roGFP2-Tpx1.C169S is partially oxidized in the cytosol of wild-type cells while HyPer7 is fully reduced. We have successfully expressed HyPer7 in the mitochondrial matrix, and it is ~40% oxidized, suggesting higher steady-state levels of peroxides, in the low micromolar range, than in the cytosol. Cytosolic HyPer7 can detect negligible H2O2 in the cytosol from mitochondrial origin unless the main H2O2 scavenger, the cytosolic peroxiredoxin Tpx1, is absent, while mitochondrial HyPer7 is oxidized to the same extent in wild-type and ∆tpx1 cells. We conclude that there is a bidirectional flux of H2O2 across the matrix and the cytosol, but Tpx1 rapidly and efficiently scavenges mitochondrial-generated peroxides and stops their steady-state cytosolic levels rising.
publishDate 2021
dc.date.none.fl_str_mv 2021
2021
2021
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10230/47747
http://dx.doi.org/10.3390/antiox10050731
url http://hdl.handle.net/10230/47747
http://dx.doi.org/10.3390/antiox10050731
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Antioxidants (Basel). 2021;10(5):731
info:eu-repo/grantAgreement/ES/2PE/PGC2018-093920-B-I00
info:eu-repo/grantAgreement/ES/2PE/PGC2018-097248-B-I00
dc.rights.none.fl_str_mv https://creativecommons.org/licenses/by/4.0/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv MDPI
publisher.none.fl_str_mv MDPI
dc.source.none.fl_str_mv reponame:Recercat. Dipósit de la Recerca de Catalunya
instname:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
instname_str Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
reponame_str Recercat. Dipósit de la Recerca de Catalunya
collection Recercat. Dipósit de la Recerca de Catalunya
repository.name.fl_str_mv
repository.mail.fl_str_mv
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