Imaging cytosolic NADH-NAD(+) redox state with a genetically encoded fluorescent biosensor.

@article{Hung2011ImagingCN,
  title={Imaging cytosolic NADH-NAD(+) redox state with a genetically encoded fluorescent biosensor.},
  author={Yin Pun Hung and John G Albeck and Mathew Tantama and Gary Yellen},
  journal={Cell metabolism},
  year={2011},
  volume={14 4},
  pages={545-54}
}
NADH is a key metabolic cofactor whose sensitive and specific detection in the cytosol of live cells has been difficult. We constructed a fluorescent biosensor of the cytosolic NADH-NAD(+) redox state by combining a circularly permuted GFP T-Sapphire with a bacterial NADH-binding protein, Rex. Although the initial construct reported [NADH] × [H(+)] / [NAD(+)], its pH sensitivity was eliminated by mutagenesis. The engineered biosensor Peredox reports cytosolic NADH:NAD(+) ratios and can be… CONTINUE READING
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