A family of NADPH/NADP+ biosensors reveals in vivo dynamics of central redox metabolism across eukaryotes

Abstract The NADPH/NADP+ redox couple is central to metabolism and redox signalling. NADP redox state is differentially regulated by distinct enzymatic machineries at the subcellular compartment level. Nonetheless, a detailed understanding of subcellular NADP redox dynamics is limited by the availab...

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Main Authors: Marie Scherschel, Jan-Ole Niemeier, Lianne J. H. C. Jacobs, Markus D. A. Hoffmann, Anika Diederich, Christopher Bell, Pascal Höhne, Sonja Raetz, Johanna B. Kroll, Janina Steinbeck, Sophie Lichtenauer, Jan Multhoff, Jannik Zimmermann, Tanmay Sadhanasatish, R. Alexander Rothemann, Carsten Grashoff, Joris Messens, Emmanuel Ampofo, Matthias W. Laschke, Jan Riemer, Leticia Prates Roma, Markus Schwarzländer, Bruce Morgan
Format: Article
Language:English
Published: Nature Portfolio 2024-12-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-024-55302-x
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author Marie Scherschel
Jan-Ole Niemeier
Lianne J. H. C. Jacobs
Markus D. A. Hoffmann
Anika Diederich
Christopher Bell
Pascal Höhne
Sonja Raetz
Johanna B. Kroll
Janina Steinbeck
Sophie Lichtenauer
Jan Multhoff
Jannik Zimmermann
Tanmay Sadhanasatish
R. Alexander Rothemann
Carsten Grashoff
Joris Messens
Emmanuel Ampofo
Matthias W. Laschke
Jan Riemer
Leticia Prates Roma
Markus Schwarzländer
Bruce Morgan
author_facet Marie Scherschel
Jan-Ole Niemeier
Lianne J. H. C. Jacobs
Markus D. A. Hoffmann
Anika Diederich
Christopher Bell
Pascal Höhne
Sonja Raetz
Johanna B. Kroll
Janina Steinbeck
Sophie Lichtenauer
Jan Multhoff
Jannik Zimmermann
Tanmay Sadhanasatish
R. Alexander Rothemann
Carsten Grashoff
Joris Messens
Emmanuel Ampofo
Matthias W. Laschke
Jan Riemer
Leticia Prates Roma
Markus Schwarzländer
Bruce Morgan
author_sort Marie Scherschel
collection DOAJ
description Abstract The NADPH/NADP+ redox couple is central to metabolism and redox signalling. NADP redox state is differentially regulated by distinct enzymatic machineries at the subcellular compartment level. Nonetheless, a detailed understanding of subcellular NADP redox dynamics is limited by the availability of appropriate tools. Here, we introduce NAPstars, a family of genetically encoded, fluorescent protein-based NADP redox state biosensors. NAPstars offer real-time, specific measurements, across a broad-range of NADP redox states, with subcellular resolution. NAPstar measurements in yeast, plants, and mammalian cell models, reveal a conserved robustness of cytosolic NADP redox homoeostasis. NAPstars uncover cell cycle-linked NADP redox oscillations in yeast and illumination- and hypoxia-dependent NADP redox changes in plant leaves. By applying NAPstars in combination with selective impairment of the glutathione and thioredoxin antioxidative pathways under acute oxidative challenge, we find an unexpected and conserved role for the glutathione system as the primary mediator of antioxidative electron flux.
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spelling doaj-art-2015d35a69e64058ba3b032da1d010e72025-08-20T01:57:16ZengNature PortfolioNature Communications2041-17232024-12-0115112010.1038/s41467-024-55302-xA family of NADPH/NADP+ biosensors reveals in vivo dynamics of central redox metabolism across eukaryotesMarie Scherschel0Jan-Ole Niemeier1Lianne J. H. C. Jacobs2Markus D. A. Hoffmann3Anika Diederich4Christopher Bell5Pascal Höhne6Sonja Raetz7Johanna B. Kroll8Janina Steinbeck9Sophie Lichtenauer10Jan Multhoff11Jannik Zimmermann12Tanmay Sadhanasatish13R. Alexander Rothemann14Carsten Grashoff15Joris Messens16Emmanuel Ampofo17Matthias W. Laschke18Jan Riemer19Leticia Prates Roma20Markus Schwarzländer21Bruce Morgan22Institute of Biochemistry, Center for Human and Molecular Biology (ZHMB), Saarland UniversityInstitute of Plant Biology and Biotechnology, University of Münster, Schlossplatz 8Redox Metabolism, Institute for Biochemistry, University of CologneDepartment of Biophysics, Center for Human and Molecular Biology (ZHMB), Saarland UniversityInstitute of Biochemistry, Center for Human and Molecular Biology (ZHMB), Saarland UniversityInstitute of Plant Biology and Biotechnology, University of Münster, Schlossplatz 8Institute of Biochemistry, Center for Human and Molecular Biology (ZHMB), Saarland UniversityInstitute of Plant Biology and Biotechnology, University of Münster, Schlossplatz 8Institute of Plant Biology and Biotechnology, University of Münster, Schlossplatz 8Institute of Plant Biology and Biotechnology, University of Münster, Schlossplatz 8Institute of Plant Biology and Biotechnology, University of Münster, Schlossplatz 8Institute of Plant Biology and Biotechnology, University of Münster, Schlossplatz 8Institute of Biochemistry, Center for Human and Molecular Biology (ZHMB), Saarland UniversityInstitute of Integrative Cell Biology and Physiology, University of Münster, Schlossplatz 5Redox Metabolism, Institute for Biochemistry, University of CologneInstitute of Integrative Cell Biology and Physiology, University of Münster, Schlossplatz 5VIB-VUB Center for Structural Biology, Vlaams Instituut voor BiotechnologieInstitute for Clinical & Experimental Surgery, Saarland UniversityInstitute for Clinical & Experimental Surgery, Saarland UniversityRedox Metabolism, Institute for Biochemistry, University of CologneDepartment of Biophysics, Center for Human and Molecular Biology (ZHMB), Saarland UniversityInstitute of Plant Biology and Biotechnology, University of Münster, Schlossplatz 8Institute of Biochemistry, Center for Human and Molecular Biology (ZHMB), Saarland UniversityAbstract The NADPH/NADP+ redox couple is central to metabolism and redox signalling. NADP redox state is differentially regulated by distinct enzymatic machineries at the subcellular compartment level. Nonetheless, a detailed understanding of subcellular NADP redox dynamics is limited by the availability of appropriate tools. Here, we introduce NAPstars, a family of genetically encoded, fluorescent protein-based NADP redox state biosensors. NAPstars offer real-time, specific measurements, across a broad-range of NADP redox states, with subcellular resolution. NAPstar measurements in yeast, plants, and mammalian cell models, reveal a conserved robustness of cytosolic NADP redox homoeostasis. NAPstars uncover cell cycle-linked NADP redox oscillations in yeast and illumination- and hypoxia-dependent NADP redox changes in plant leaves. By applying NAPstars in combination with selective impairment of the glutathione and thioredoxin antioxidative pathways under acute oxidative challenge, we find an unexpected and conserved role for the glutathione system as the primary mediator of antioxidative electron flux.https://doi.org/10.1038/s41467-024-55302-x
spellingShingle Marie Scherschel
Jan-Ole Niemeier
Lianne J. H. C. Jacobs
Markus D. A. Hoffmann
Anika Diederich
Christopher Bell
Pascal Höhne
Sonja Raetz
Johanna B. Kroll
Janina Steinbeck
Sophie Lichtenauer
Jan Multhoff
Jannik Zimmermann
Tanmay Sadhanasatish
R. Alexander Rothemann
Carsten Grashoff
Joris Messens
Emmanuel Ampofo
Matthias W. Laschke
Jan Riemer
Leticia Prates Roma
Markus Schwarzländer
Bruce Morgan
A family of NADPH/NADP+ biosensors reveals in vivo dynamics of central redox metabolism across eukaryotes
Nature Communications
title A family of NADPH/NADP+ biosensors reveals in vivo dynamics of central redox metabolism across eukaryotes
title_full A family of NADPH/NADP+ biosensors reveals in vivo dynamics of central redox metabolism across eukaryotes
title_fullStr A family of NADPH/NADP+ biosensors reveals in vivo dynamics of central redox metabolism across eukaryotes
title_full_unstemmed A family of NADPH/NADP+ biosensors reveals in vivo dynamics of central redox metabolism across eukaryotes
title_short A family of NADPH/NADP+ biosensors reveals in vivo dynamics of central redox metabolism across eukaryotes
title_sort family of nadph nadp biosensors reveals in vivo dynamics of central redox metabolism across eukaryotes
url https://doi.org/10.1038/s41467-024-55302-x
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