文献类型：期刊文献

英文题名：Genetically encoded fluorescent sensors reveal dynamic regulation of NADPH metabolism

作者：Tao, Rongkun[1];Zhao, Yuzheng[1,2,3];Chu, Huanyu[4];Wang, Aoxue[1];Zhu, Jiahuan[1];Chen, Xianjun[1,2,3];Zou, Yejun[1];Shi, Mei[1];Liu, Renmei[1];Su, Ni[1];Du, Jiulin[2];Zhou, Hai-Meng[5];Zhu, Linyong[6];Qian, Xuhong[7];Liu, Haiyan[4];Loscalzo, Joseph[8];Yang, Yi[1,2]

机构：[1]East China Univ Sci & Technol, Shanghai Collaborat Innovat Ctr Biomfg Technol, Synthet Biol & Biotechnol Lab, State Key Lab Bioreactor Engn, Shanghai, Peoples R China;[2]Chinese Acad Sci, Inst Neurosci, CAS Ctr Excellence Brain Sci, State Key Lab Neurosci, Shanghai, Peoples R China;[3]East China Univ Sci & Technol, Shanghai Key Lab New Drug Design, Sch Pharm, Shanghai, Peoples R China;[4]Univ Sci & Technol China, Hefei Natl Lab Phys Sci Microscale, Sch Life Sci, Hefei, Anhui, Peoples R China;[5]Tsinghua Univ, Yangtze Delta Reg Inst, Zhejiang Prov Key Lab Appl Enzymol, Jiaxing, Peoples R China;[6]East China Univ Sci & Technol, Sch Chem & Mol Engn, Shanghai, Peoples R China;[7]East China Univ Sci & Technol, Shanghai Key Lab Chem Biol, Sch Pharm, Shanghai, Peoples R China;[8]Harvard Med Sch, Brigham & Womens Hosp, Dept Med, Boston, MA USA

年份：2017

卷号：14

期号：7

起止页码：720

外文期刊名：NATURE METHODS

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000404399500030)】；

基金：We thank S.J. Remington for the roGFP1 vector; G. Yellen for the PercevalHR vector, V.V. Belousov for the HyPerRed vector; C. Fan, J. Ye, Y. Tang, T. Wang, Z. Wu, H. Hu, Z. Du, T. Li, Y. Xia, L. Zhou and Y. Wang for technical assistance; and S. Tribune for secretarial assistance. This research was supported by the National Natural Science Foundation of China (31225008, 91313301 and 31470833 to Y.Y. 31370755 to H.L.; 91649123 and 31671484 to Y. Zhao), 973 Program (2013CB531200 to Y.Y.), the Shanghai Science and Technology Commission (14XD1401400 and 16430723100 to Y.Y. and 15YF1402600 to Y. Zhao), the Specialized Research Fund for the Doctoral Program of Higher Education (20100074110010 to Y.Y.), the Lift Engineering for Young Talent of China Association for Science and Technology (to Y. Zhao), Shanghai Young Topnotch Talent (to Y. Zhao), the State Key Laboratory of Bioreactor Engineering (to Y.Y.), the 111 Project (607023 to Y.Y.), the Fundamental Research Funds for the Central Universities (to Y.Y. and Y. Zhao) and the US National Institutes of Health (HL061795, HL007690 and GM107618 to J.L.).

语种：英文

摘要：Reduced nicotinamide adenine dinucleotide phosphate (NADPH) is essential for biosynthetic reactions and antioxidant functions; however, detection of NADPH metabolism in living cells remains technically challenging. We develop and characterize ratiometric, pH-resistant, genetically encoded fluorescent indicators for NADPH (iNap sensors) with various affinities and wide dynamic range. iNap sensors enabled quantification of cytosolic and mitochondria( NADPH pools that are controlled by cytosolic NAD+ kinase levels and revealed cellular NADPH dynamics under oxidative stress depending on glucose availability. We found that mammalian cells have a strong tendency to maintain physiological NADPH homeostasis, which is regulated by glucose-6-phosphate dehydrogenase and AMP kinase. Moreover, using the iNap sensors we monitor NADPH fluctuations during the activation of macrophage cells or wound response in vivo. These data demonstrate that the iNap sensors will be valuable tools for monitoring NADPH dynamics in live cells and gaining new insights into cell metabolism.