A genetically encoded tool to increase cellular NADH/NAD + ratio in living cells.

Autor: Pan X; Laboratory of Redox Biology and Metabolism, Scintillon Institute, San Diego, CA, USA., Heacock ML; Laboratory of Redox Biology and Metabolism, Scintillon Institute, San Diego, CA, USA.; Calibr, The Scripps Research Institute, La Jolla, CA, USA., Abdulaziz EN; Laboratory of Redox Biology and Metabolism, Scintillon Institute, San Diego, CA, USA.; Process Development Associate, Amgen, Thousand Oaks, CA, USA., Violante S; Donald B. and Catherine C. Marron Cancer Metabolism Center, Memorial Sloan Kettering Cancer Center, New York, NY, USA., Zuckerman AL; Laboratory of Redox Biology and Metabolism, Scintillon Institute, San Diego, CA, USA.; Program in Mathematics and Science Education, University of California San Diego, San Diego, CA, USA.; Program in Mathematics and Science Education, San Diego State University, San Diego, USA., Shrestha N; Liver Center, Division of Gastroenterology, Massachusetts General Hospital, Boston, MA, USA., Yao C; Laboratory of Redox Biology and Metabolism, Scintillon Institute, San Diego, CA, USA., Goodman RP; Liver Center, Division of Gastroenterology, Massachusetts General Hospital, Boston, MA, USA., Cross JR; Donald B. and Catherine C. Marron Cancer Metabolism Center, Memorial Sloan Kettering Cancer Center, New York, NY, USA., Cracan V; Laboratory of Redox Biology and Metabolism, Scintillon Institute, San Diego, CA, USA. vcracan@scripps.edu.; Department of Chemistry, The Scripps Research Institute, La Jolla, CA, USA. vcracan@scripps.edu.
Jazyk: angličtina
Zdroj: Nature chemical biology [Nat Chem Biol] 2024 May; Vol. 20 (5), pp. 594-604. Date of Electronic Publication: 2023 Oct 26.
DOI: 10.1038/s41589-023-01460-w
Abstrakt: 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.
(© 2023. The Author(s), under exclusive licence to Springer Nature America, Inc.)
Databáze: MEDLINE
Externí odkaz: