1. Biochemistry and Chemical Biology
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Semisynthetic biosensors for mapping cellular concentrations of nicotinamide adenine dinucleotides

  1. Olivier Sallin
  2. Luc Reymond
  3. Corentin Gondrand
  4. Fabio Raith
  5. Birgit Koch
  6. Kai Johnsson  Is a corresponding author
  1. École Polytechnique Fédérale de Lausanne, Switzerland
  2. Max Planck Institute for Medical Research, Germany
  3. Max-Planck-Institute of Medical Research, Germany
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Cite this article as: eLife 2018;7:e32638 doi: 10.7554/eLife.32638

Abstract

We introduce a new class of semisynthetic fluorescent biosensors for the quantification of free nicotinamide adenine dinucleotide (NAD+) and ratios of reduced to oxidized nicotinamide adenine dinucleotide phosphate (NADPH/NADP+) in live cells. Sensing is based on controlling the spatial proximity of two synthetic fluorophores by binding of NAD(P) to the protein component of the sensor. The sensors possess a large dynamic range, can be excited at long wavelengths, are pH-insensitive, have tunable response range and can be localized in different organelles. Ratios of free NADPH/NADP+ are found to be higher in mitochondria compared to those found in the nucleus and the cytosol. By recording free NADPH/NADP+ ratios in response to changes in environmental conditions, we observe how cells can react to such changes by adapting metabolic fluxes. Finally, we demonstrate how a comparison of the effect of drugs on cellular NAD(P) levels can be used to probe mechanisms of action.

Article and author information

Author details

  1. Olivier Sallin

    Institute of Chemical Sciences and Engineering, École Polytechnique Fédérale de Lausanne, Lausanne, Switzerland
    Competing interests
    Olivier Sallin, has filed a patent application (WO2016131833A1) on the design and use of sensors for the detection of NAD(P).
  2. Luc Reymond

    Institute of Chemical Sciences and Engineering, École Polytechnique Fédérale de Lausanne, Lausanne, Switzerland
    Competing interests
    Luc Reymond, has filed a patent application (WO2016131833A1) on the design and use of sensors for the detection of NAD(P).
  3. Corentin Gondrand

    Department of Chemical Biology, Max Planck Institute for Medical Research, Heidelberg, Germany
    Competing interests
    No competing interests declared.
  4. Fabio Raith

    Department of Chemical Biology, Max Planck Institute for Medical Research, Heidelberg, Germany
    Competing interests
    No competing interests declared.
  5. Birgit Koch

    Department of Chemical Biology, Max-Planck-Institute of Medical Research, Heidelberg, Germany
    Competing interests
    No competing interests declared.
  6. Kai Johnsson

    Institute of Chemical Sciences and Engineering, École Polytechnique Fédérale de Lausanne, Lausanne, Switzerland
    For correspondence
    kai.johnsson@epfl.ch
    Competing interests
    Kai Johnsson, has filed a patent application (WO2016131833A1) on the design and use of sensors for the detection of NAD(P).
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8002-1981

Funding

Max-Planck-Gesellschaft (Institutional support and open-access funding)

  • Kai Johnsson

Ecole Polytechnique Federale de Lausanne (Institutional support and open-access funding)

  • Corentin Gondrand

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Yamuna Krishnan, University of Chicago, United States

Publication history

  1. Received: October 9, 2017
  2. Accepted: May 9, 2018
  3. Accepted Manuscript published: May 29, 2018 (version 1)
  4. Version of Record published: June 6, 2018 (version 2)

Copyright

© 2018, Sallin et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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