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NERNST: a genetically-encoded ratiometric non-destructive sensing tool to estimate NADP(H) redox status in bacterial, plant and animal systems

Author

Listed:
  • Pamela E. Molinari

    (Universidad Nacional de Rosario (UNR))

  • Adriana R. Krapp

    (Universidad Nacional de Rosario (UNR))

  • Andrea Weiner

    (Universidad Nacional de Rosario (UNR))

  • Hannes M. Beyer

    (University of Düsseldorf)

  • Arun Kumar Kondadi

    (Medical Faculty and University Hospital Düsseldorf, Heinrich-Heine-University Düsseldorf)

  • Tim Blomeier

    (University of Düsseldorf)

  • Melina López

    (Universidad Nacional de Rosario (UNR))

  • Pilar Bustos-Sanmamed

    (Universidad Nacional de Rosario (UNR))

  • Evelyn Tevere

    (Universidad Nacional de Rosario (UNR))

  • Wilfried Weber

    (University of Freiburg
    Saarland University)

  • Andreas S. Reichert

    (Medical Faculty and University Hospital Düsseldorf, Heinrich-Heine-University Düsseldorf)

  • Nora B. Calcaterra

    (Universidad Nacional de Rosario (UNR))

  • Mathias Beller

    (University of Düsseldorf)

  • Nestor Carrillo

    (Universidad Nacional de Rosario (UNR))

  • Matias D. Zurbriggen

    (University of Düsseldorf
    CEPLAS – Cluster of Excellence on Plant Sciences)

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.

Suggested Citation

  • Pamela E. Molinari & Adriana R. Krapp & Andrea Weiner & Hannes M. Beyer & Arun Kumar Kondadi & Tim Blomeier & Melina López & Pilar Bustos-Sanmamed & Evelyn Tevere & Wilfried Weber & Andreas S. Reicher, 2023. "NERNST: a genetically-encoded ratiometric non-destructive sensing tool to estimate NADP(H) redox status in bacterial, plant and animal systems," Nature Communications, Nature, vol. 14(1), pages 1-18, December.
    • Handle: RePEc:nat:natcom:v:14:y:2023:i:1:d:10.1038_s41467-023-38739-4
    DOI: 10.1038/s41467-023-38739-4
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    References listed on IDEAS

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    1. Yulia G. Ermakova & Dmitry S. Bilan & Mikhail E. Matlashov & Natalia M. Mishina & Ksenia N. Markvicheva & Oksana M. Subach & Fedor V. Subach & Ivan Bogeski & Markus Hoth & Grigori Enikolopov & Vsevolo, 2014. "Red fluorescent genetically encoded indicator for intracellular hydrogen peroxide," Nature Communications, Nature, vol. 5(1), pages 1-9, December.
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