A genetically encoded tool to increase cellular NADH/NAD(+) ratio in living cells Journal Article
| Authors: | Pan, X.; Heacock, M. L.; Abdulaziz, E. N.; Violante, S.; Zuckerman, A. L.; Shrestha, N.; Yao, C.; Goodman, R. P.; Cross, J. R.; Cracan, V. |
| Article Title: | A genetically encoded tool to increase cellular NADH/NAD(+) ratio in living cells |
| Abstract: | Impaired redox metabolism is a key contributor to the etiology of many diseases, including primary mitochondrial disorders, cancer, neurodegeneration and aging. However, mechanistic studies of redox imbalance remain challenging due to limited strategies that can perturb redox metabolism in various cellular or organismal backgrounds. Most studies involving impaired redox metabolism have focused on oxidative stress; consequently, less is known about the settings where there is an overabundance of NADH reducing equivalents, termed reductive stress. Here we introduce a soluble transhydrogenase from Escherichia coli (EcSTH) as a novel genetically encoded tool to promote reductive stress in living cells. When expressed in mammalian cells, EcSTH, and a mitochondrially targeted version (mitoEcSTH), robustly elevated the NADH/NAD+ ratio in a compartment-specific manner. Using this tool, we determined that metabolic and transcriptomic signatures of the NADH reductive stress are cellular background specific. Collectively, our novel genetically encoded tool represents an orthogonal strategy to promote reductive stress. (Figure presented.). © The Author(s), under exclusive licence to Springer Nature America, Inc. 2023. |
| Keywords: | controlled study; unclassified drug; human cell; promoter region; genetics; nonhuman; cell proliferation; metabolism; serine; carboxy terminal sequence; transcriptomics; escherichia coli; western blotting; citric acid; cellular distribution; fluorescence microscopy; down regulation; lactate dehydrogenase; oxidative stress; growth differentiation factor 15; superoxide; enzyme specificity; mitochondria; nicotinamide adenine dinucleotide (phosphate) transhydrogenase; oxygen consumption; genetic code; mitochondrion; glycolysis; aspartic acid; reduced nicotinamide adenine dinucleotide; oxidation reduction reaction; nad; oxidation-reduction; absorption spectroscopy; pseudomonas putida; glutathione; cell stress; escherichia coli proteins; nicotinamide adenine dinucleotide; mammal cell; pyruvic acid; citric acid cycle; escherichia coli protein; metabolomics; hek293 cells; liquid chromatography-mass spectrometry; pentose phosphate cycle; malic acid; glutathione disulfide; azotobacter vinelandii; succinic acid; oligomycin; whole cell; ribose 5 phosphate; humans; human; female; article; hela cell line; oxidation reduction potential; live cell imaging; acetoacetic acid; glucose 6 phosphate; 3 hydroxybutyric acid; hek293t cell line; antimycin a1; a-549 cell line; metabolic fingerprinting; panc-1 cell line; fructose 1 phosphate; fetal bovine serum; hek293s cell line; dihydroxyacetone phosphate; fructose 1,6 bisphosphate; fructose 6 phosphate; fumaric acid; nadp transhydrogenases; 3 phosphoglycerate; 6 phosphogluconate; erythrose 4 phosphate; n acetylglucosamine 1 phosphate; sedoheptulose 7 phosphate; xylulose 5 phosphate; c2c12 cell line; cuniculidae; imr-90 cell line; metabolic ratio |
| Journal Title: | Nature Chemical Biology |
| Volume: | 20 |
| Issue: | 5 |
| ISSN: | 1552-4450 |
| Publisher: | Nature Publishing Group |
| Date Published: | 2024-05-01 |
| Start Page: | 594 |
| End Page: | 604 |
| Language: | English |
| DOI: | 10.1038/s41589-023-01460-w |
| PUBMED: | 37884806 |
| PROVIDER: | scopus |
| PMCID: | PMC11045668 |
| DOI/URL: | |
| Notes: | Article -- Source: Scopus |
