title: Genetically encoded fluorescent sensors for intracellular NADH detection.
abstract: We have developed genetically encoded fluorescent sensors for reduced nicotinamide adenine dinucleotide (NADH), which manifest a large change in fluorescence upon NADH binding. We demonstrate the utility of these sensors in mammalian cells by monitoring the dynamic changes in NADH levels in subcellular organelles as affected by NADH transport, glucose metabolism, electron transport chain function, and redox environment, and we demonstrate the temporal separation of changes in mitochondrial and cytosolic NADH levels with perturbation. These results support the view that cytosolic NADH is sensitive to environmental changes, while mitochondria have a strong tendency to maintain physiological NADH homeostasis. These sensors provide a very good alternative to existing techniques that measure endogenous fluorescence of intracellular NAD(P)H and, owing to their superior sensitivity and specificity, allow for the selective monitoring of total cellular and compartmental responses of this essential cofactor.
authors: Synthetic Biology and Biotechnology Laboratory, State Key Laboratory of Bioreactor Engineering, School of pharmacy, East China University of Science and Technology, Shanghai 200237, China.
authors: Yuzheng
authors: Y
authors: Zhao
authors: Jing
authors: J
authors: Jin
authors: Qingxun
authors: Q
authors: Hu
authors: Hai-Meng
authors: HM
authors: Zhou
authors: Jing
authors: J
authors: Yi
authors: Zhenhang
authors: Z
authors: Yu
authors: Lei
authors: L
authors: Xu
authors: Xue
authors: X
authors: Wang
authors: Yi
authors: Y
authors: Yang
authors: Joseph
authors: J
authors: Loscalzo
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D003600:Cytosol / Q000378:metabolism; D005947:Glucose / Q000494:pharmacology; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D019344:Lactic Acid / Q000494:pharmacology; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D009243:NAD / Q000032:analysis*; D010084:Oxidation-Reduction; D019289:Pyruvic Acid / Q000494:pharmacology
chemical_list: D001426:Bacterial Proteins; D008164:Luminescent Proteins; C054562:yellow fluorescent protein, Bacteria; D009243:NAD; D019344:Lactic Acid; D019289:Pyruvic Acid; D006861:Hydrogen Peroxide; D005947:Glucose