School of Chemistry and Chemical Engineering, Yan'an University, Yan'an 716000, P. R. China.
Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province, School of Chemistry and Chemical Engineering, Shaanxi Normal University, Xi'an, 710062, P. R. China.
Anal Chem. 2020 Aug 18;92(16):10940-10945. doi: 10.1021/acs.analchem.0c00569. Epub 2020 Aug 6.
Monitoring the fluctuation of adenosine 5'-triphosphate (ATP) at the subcellular level is important for the study of cell energy metabolism. Herein, we fabricated an electrochemical nanoaptasensor for continuously monitoring ATP fluctuation at the subcellular level. A gold nanoelectrode with a diameter of 120 nm was fabricated, and ferrocene (Fc)-labeled anti-ATP aptamer was self-assembled onto the nanoelectrode surface to form a nanoaptasensor. In the presence of ATP, the ferrocene-labeled anti-ATP aptamer bound with two ATP units to form an ATP-aptamer conjugation, resulting in the close proximity of Fc to the nanoelectrode surface and then an increase of oxidation current of Fc. ATP can be detected with a detection limit of 26 μM within 2 min. Cell viability assays indicated that the nanoaptasensor was biocompatible with negligible biological effects. By taking advantage of the good biocompatibility of the nanoaptasensor, ATP fluctuation at the subcellular level was monitored under glucose starvation and Ca induction. This work demonstrates that the nanoaptasensor is a useful tool for investigating ATP-relevant biological processes via the electrochemical method.
监测细胞能量代谢中三磷酸腺苷（ATP）在亚细胞水平的波动非常重要。在此，我们构建了一种电化学纳米适体传感器，用于连续监测亚细胞水平的 ATP 波动。制备了直径为 120nm 的金纳米电极，并将二茂铁（Fc）标记的抗-ATP 适体自组装到纳米电极表面，形成纳米适体传感器。在存在 ATP 的情况下，Fc 标记的抗-ATP 适体与两个 ATP 单元结合形成 ATP-适体缀合物，导致 Fc 与纳米电极表面接近，从而增加 Fc 的氧化电流。在 2 分钟内可以检测到检测限为 26μM 的 ATP。细胞活力测定表明，纳米适体传感器具有良好的生物相容性，几乎没有生物学效应。利用纳米适体传感器的良好生物相容性，在葡萄糖饥饿和 Ca 诱导下监测了亚细胞水平的 ATP 波动。这项工作表明，该纳米适体传感器是通过电化学方法研究与 ATP 相关的生物学过程的有用工具。
