A family of NADPH/NADP+ biosensors reveals in vivo dynamics of central redox metabolism across eukaryotes
The NADPH/NADP<sup>+</sup> 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.
Authors
- Emmanuel Ampofo
- S. Raetz (ORCID: https://orcid.org/0009-0000-0896-8700)
- Letícia Prates Roma (ORCID: https://orcid.org/0000-0001-9527-0529)
- Jan Riemer (ORCID: https://orcid.org/0000-0002-7574-8457)
- Robin Alexander Rothemann (ORCID: https://orcid.org/0000-0002-9502-5360)
- Matthias W. Laschke (ORCID: https://orcid.org/0000-0002-7847-8456)
- Tanmay Sadhanasatish (ORCID: https://orcid.org/0000-0003-4522-4115)
- Johanna B. Kroll (ORCID: https://orcid.org/0000-0003-4652-3598)
- Lianne H.C. Jacobs (ORCID: https://orcid.org/0000-0001-5090-601X)
- Jan Multhoff (ORCID: https://orcid.org/0009-0005-3409-461X)
- Markus Schwarzländer (ORCID: https://orcid.org/0000-0003-0796-8308)
- Carsten Grashoff (ORCID: https://orcid.org/0000-0003-0118-9231)
- Jan-Ole Niemeier (ORCID: https://orcid.org/0000-0002-4081-8101)
- Christopher Bell (ORCID: https://orcid.org/0009-0009-4689-9035)
- Janina Steinbeck (ORCID: https://orcid.org/0000-0002-7793-5053)
- Bruce Morgan (ORCID: https://orcid.org/0000-0001-9393-1071)
- Sophie Lichtenauer (ORCID: https://orcid.org/0000-0003-4525-6555)
- Joris Messens (ORCID: https://orcid.org/0000-0002-2128-8264)
- Markus D. A. Hoffmann (ORCID: https://orcid.org/0000-0001-6765-3469)
- Jannik Zimmermann (ORCID: https://orcid.org/0000-0002-3336-042X)
- Pascal Höhne (ORCID: https://orcid.org/0009-0007-1128-2110)
- Marie Scherschel (ORCID: https://orcid.org/0000-0003-1907-7852)
- Anika Diederich (ORCID: https://orcid.org/0009-0001-6234-4744)
Institutions
- Vrije Universiteit Brussel (BE)
- University of Cologne (DE)
- University of Münster (DE)
- Vlaams Instituut voor Biotechnologie (BE)
- Institute of Plant Biology and Biotechnology (KZ)
- Cologne Excellence Cluster on Cellular Stress Responses in Aging Associated Diseases (DE)
- VIB-VUB Center for Structural Biology (BE)
- Saarland University (DE)
Publication Details
- Journal
- Nature Communications
- Published
- 2024-12-19
- DOI
- https://doi.org/10.1038/s41467-024-55302-x
- Citations
- 30
- Primary Topic
- Redox biology and oxidative stress
- Type
- article
- Field-Weighted Citation Impact
- 7.01
Funders
- Deutsche Forschungsgemeinschaft
- Universität des Saarlandes