| Title | Live-Cell Imaging of NADPH Production from Specific Pathways |
| Authors | Hong, Senlian Chen, Tao Liu, Ling Cao, Chen Lv, Fengxiang Rabinowitz, Joshua D. Huang, Yanyi Chen, Xing |
| Affiliation | Peking Univ, Coll Chem & Mol Engn, Beijing 100871, Peoples R China Peking Univ, Peking Tsinghua Ctr Life Sci, Beijing 100871, Peoples R China Peking Univ, Beijing Natl Lab Mol Sci, Beijing 100871, Peoples R China Peking Univ, Coll Engn, Beijing 100871, Peoples R China Peking Univ, Biomed Pioneering Innovat Ctr, Beijing 100871, Peoples R China Peking Univ, Beijing Adv Innovat Ctr Genom, Beijing 100871, Peoples R China Princeton Univ, Lewis Sigler Inst Integrat Genom, Princeton, NJ 08544 USA Princeton Univ, Dept Chem, Princeton, NJ 08544 USA Peking Univ, Inst Mol Med, Beijing 100871, Peoples R China Peking Univ, Inst Cell Anal, Shenzhen Bey Lab, Beijing 100871, Peoples R China Peking Univ, Synthet & Funct Biomol Ctr, Beijing 100871, Peoples R China Peking Univ, Key Lab Bioorgan Chem & Mol Engn, Minist Educ, Beijing 100871, Peoples R China Scripps Res Inst, Dept Mol Med, La Jolla, CA 92037 USA Baylor Coll Med, Huffington Ctr Aging, Houston, TX 77030 USA |
| Keywords | MALIC ENZYME |
| Issue Date | 2021 |
| Publisher | CCS CHEMISTRY |
| Abstract | As anconversion of essential cofactor for lipid bio-synthesis and antioxidant defense, reduced nicotinamide adenine dinucleotide phosphate (NADPH) is produced via various pathways, including the oxidative pentose phosphate pathway (oxPPP) and the malic enzyme 1 (ME1)-catalyzed conversion of malate to pyruvate. Live-cell detection of NADPH production routes remains challenging. Here, we report tracing hydrides into lipid droplets (THILD), a chemical imaging strategy for the detection of pathway-specific NADPH generation in live cells. This strategy exploits deuterium (2H)-labeled glucose ([2H]Glc) tracers that transfer deuterides to NADPH via specific pathways. The NADP(2)H, in turn, transfers deuterides to lipids, resulting in accumulation of C-H-2 bonds in lipid droplets, which can be visualized by bioorthogonal stimulated Raman scattering (SRS) microscopy. We used this concept to demonstrate the imaging of oxPPP-produced NADPH using the oxPPP-specific tracer, [3-H-2]Glc. Furthermore, the "switch on" of NADPH production by ME1 in differentiating adipocytes was imaged by [4-H-2]Glc. Finally, comparison of [3-2H]Glc and [4-2H]Glc THILD imaging of adipocytes showed that hypoxia induces suppression of ME1-mediated NADPH production and oxPPP-produced NADPH becomes the main source. |
| URI | http://hdl.handle.net/20.500.11897/643365 |
| DOI | 10.31635/ccschem.020.202000346 |
| Indexed | ESCI |
| Appears in Collections: | å å¦ä¸ å å å·¥ç¨ å¦é ¢ 生命科学学院 工学院 分子医学研究所 生物有机与分子工程教育部重点实验室 |
