PUBLICATION
NERNST: a genetically-encoded ratiometric non-destructive sensing tool to estimate NADP(H) redox status in bacterial, plant and animal systems
- Authors
- Molinari, P.E., Krapp, A.R., Weiner, A., Beyer, H.M., Kondadi, A.K., Blomeier, T., López, M., Bustos-Sanmamed, P., Tevere, E., Weber, W., Reichert, A.S., Calcaterra, N.B., Beller, M., Carrillo, N., Zurbriggen, M.D.
- ID
- ZDB-PUB-230608-29
- Date
- 2023
- Source
- Nature communications 14: 32773277 (Journal)
- Registered Authors
- Calcaterra, Nora
- Keywords
- none
- MeSH Terms
-
- Animals
- Chloroplasts/metabolism
- Mammals/metabolism
- NADP/metabolism
- Oxidation-Reduction
- Plants*/genetics
- Plants*/metabolism
- Zebrafish*/metabolism
- PubMed
- 37280202 Full text @ Nat. Commun.
Citation
Molinari, P.E., Krapp, A.R., Weiner, A., Beyer, H.M., Kondadi, A.K., Blomeier, T., López, M., Bustos-Sanmamed, P., Tevere, E., Weber, W., Reichert, A.S., Calcaterra, N.B., Beller, M., Carrillo, N., Zurbriggen, M.D. (2023) NERNST: a genetically-encoded ratiometric non-destructive sensing tool to estimate NADP(H) redox status in bacterial, plant and animal systems. Nature communications. 14:32773277.
Abstract
NADP(H) is a central metabolic hub providing reducing equivalents to multiple biosynthetic, regulatory and antioxidative pathways in all living organisms. While biosensors are available to determine NADP+ or NADPH levels in vivo, no probe exists to estimate the NADP(H) redox status, a determinant of the cell energy availability. We describe herein the design and characterization of a genetically-encoded ratiometric biosensor, termed NERNST, able to interact with NADP(H) and estimate ENADP(H). NERNST consists of a redox-sensitive green fluorescent protein (roGFP2) fused to an NADPH-thioredoxin reductase C module which selectively monitors NADP(H) redox states via oxido-reduction of the roGFP2 moiety. NERNST is functional in bacterial, plant and animal cells, and organelles such as chloroplasts and mitochondria. Using NERNST, we monitor NADP(H) dynamics during bacterial growth, environmental stresses in plants, metabolic challenges to mammalian cells, and wounding in zebrafish. NERNST estimates the NADP(H) redox poise in living organisms, with various potential applications in biochemical, biotechnological and biomedical research.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping