State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, China.
Department of Chemical Biology, Max Planck Institute for Medical Research, Jahnstrasse 29, Heidelberg 69120, Germany.
Anal Chem. 2020 Mar 17;92(6):4681-4688. doi: 10.1021/acs.analchem.0c00506. Epub 2020 Mar 5.
Real-time imaging and quantification of adenosine triphosphate (ATP) fluctuation in cells are significant for understanding the relationship between energy metabolism and cell functions. However, few synthetic fluorescent probes have been reported to tackle this challenge due to lack of accurate fluorescence readout and suitable response concentration. Herein we designed and synthesized a ratiometric fluorescent probe (Rh6G-ACFPN) for quantitatively detecting the fluctuation of mitochondrial ATP in living cells. Rh6G-ACFPN selectively and reversibly responds to ATP with an ideal dissociation constant () of 4.65 mM (3-10 mM: the range of mitochondrial ATP concentrations). Live-cell imaging allows us to directly monitor the dynamic changes of mitochondrial ATP in high temporal resolution. Moreover, for the first time, mitochondrial ATP in normal and cancer cells lines was successfully quantified and discriminated. These results demonstrate the versatility of Rh6G-ACFPN as a useful imaging tool to elucidate the function of mitochondrial ATP in living cells.
实时成像和定量检测细胞内三磷酸腺苷 (ATP) 的波动对于理解能量代谢与细胞功能之间的关系具有重要意义。然而，由于缺乏准确的荧光读出和合适的响应浓度，很少有合成荧光探针能够应对这一挑战。在此，我们设计并合成了一种比率型荧光探针（Rh6G-ACFPN），用于定量检测活细胞中线粒体 ATP 的波动。Rh6G-ACFPN 选择性和可逆地响应 ATP，其理想解离常数 () 为 4.65 mM（3-10 mM：线粒体 ATP 浓度范围）。活细胞成像使我们能够直接以高时间分辨率监测线粒体 ATP 的动态变化。此外，我们首次成功定量和区分了正常和癌细胞系中的线粒体 ATP。这些结果表明 Rh6G-ACFPN 作为一种有用的成像工具，可用于阐明线粒体 ATP 在活细胞中的功能。
