| Literature DB >> 28581494 |
Rongkun Tao1, Yuzheng Zhao1,2,3, Huanyu Chu4, Aoxue Wang1, Jiahuan Zhu1, Xianjun Chen1,2,3, Yejun Zou1, Mei Shi1, Renmei Liu1, Ni Su1, Jiulin Du2, Hai-Meng Zhou5, Linyong Zhu6, Xuhong Qian7, Haiyan Liu4, Joseph Loscalzo8, Yi Yang1,2.
Abstract
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 mitochondrial 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.Entities:
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Year: 2017 PMID： 28581494 PMCID： PMC5555402 DOI： 10.1038/nmeth.4306
Source DB: PubMed Journal: Nat Methods ISSN： 1548-7091 Impact factor: 28.547