文献类型：期刊文献

英文题名：Genetically Encoded Fluorescent Sensors for Intracellular NADH Detection

作者：Zhao, Yuzheng[1];Jin, Jing[1];Hu, Qingxun[1];Zhou, Hai-Meng[2];Yi, Jing[3];Yu, Zhenhang[4];Xu, Lei[1];Wang, Xue[1];Yang, Yi[1];Loscalzo, Joseph[5]

机构：[1]E China Univ Sci & Technol, Synthet Biol & Biotechnol Lab, State Key Lab Bioreactor Engn, Sch Pharm, Shanghai 200237, Peoples R China;[2]Tsinghua Univ, Sch Life Sci, Minist Educ, Prot Sci Lab, Beijing 100084, Peoples R China;[3]Shanghai Jiao Tong Univ, Sch Med, Dept Biochem & Mol Cell Biol,Inst Med Sci, Key Lab,Educ Minist Cell Differentiat & Apoptosis, Shanghai 200025, Peoples R China;[4]China Natl Ctr Biotechnol Dev, Beijing 100039, Peoples R China;[5]Harvard Univ, Sch Med, Brigham & Womens Hosp, Cardiovasc Div,Dept Med, Boston, MA 02115 USA

年份：2011

卷号：14

期号：4

起止页码：555

外文期刊名：CELL METABOLISM

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000296223600015)】；

基金：We thank Chunyuan Zhou, Minyan Deng, Jin Xu, and Wenyun Zheng for technical assistance and Stephanie Tribuna and Susan Vignolo-Collazzo for expert secretarial assistance. This work was supported by the NSFC (grant numbers 31071260, 90713026, 31170815, and 91013012), the 863 Program (grant number 2006AA02Z160), Specialized Research Fund for the Doctoral Program of Higher Education (grant number 20100074110010), the Fok Ying Tung Education Foundation (grant number 1110221, the Program for Professor of Special Appointment (Eastern Scholar) at Shanghai Institutions of Higher Learning, Open Funding Project of the State Key Laboratory of Bioreactor Engineering and the 111 Project (grant number B07023), and NIH grants HL 061795, HL 070819, HL 048743, HL 107192, and HL 108630 (to J.L.).

语种：英文

摘要：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.