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Journal Article

Semisynthetic biosensors for mapping cellular concentrations of nicotinamide adenine dinucleotides

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Reymond,  Luc
Max Planck Institute for Medical Research, Max Planck Society;

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Johnsson,  Kai
Chemical Biology, Max Planck Institute for Medical Research, Max Planck Society;

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Citation

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.


Cite as: http://hdl.handle.net/21.11116/0000-0001-691F-8
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.