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  Semisynthetic biosensors for mapping cellular concentrations of nicotinamide adenine dinucleotides

Sallin, O., Reymond, L., Gondrand, C., Raith, F., Koch, B., & Johnsson, K. (2018). Semisynthetic biosensors for mapping cellular concentrations of nicotinamide adenine dinucleotides. eLife, 7, 1-52. doi:10.7554/eLife.32638.

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elife_7_2018_e32638.pdf (Preprint), 2MB
 
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Sallin , Olivier, Author
Reymond, Luc1, Author              
Gondrand , Corentin, Author
Raith , Fabio, Author
Koch , Birgit, Author
Johnsson, Kai2, Author              
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1Max Planck Institute for Medical Research, Max Planck Society, ou_1125545              
2Chemical Biology, Max Planck Institute for Medical Research, Max Planck Society, ou_2364732              

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 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.

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Language(s): eng - English
 Dates: 2018-05-29
 Publication Status: Published in print
 Pages: 52
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 Table of Contents: -
 Rev. Type: Peer
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Title: eLife
Source Genre: Journal
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Publ. Info: Cambridge : eLife Sciences Publications
Pages: - Volume / Issue: 7 Sequence Number: - Start / End Page: 1 - 52 Identifier: ISSN: 2050-084X
CoNE: https://pure.mpg.de/cone/journals/resource/2050-084X