Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, Sendai, Miyagi 980-8577, Japan; Department of Molecular and Chemical Life Sciences, Graduate School of Life Sciences, Tohoku University, Sendai, Miyagi 980-8577, Japan; Department of Chemistry, Graduate School of Science, Tohoku University, Sendai, Miyagi 980-8578, Japan.
Department of Molecular and Chemical Life Sciences, Graduate School of Life Sciences, Tohoku University, Sendai, Miyagi 980-8577, Japan.
Cell Chem Biol. 2020 Dec 17;27(12):1521-1531.e8. doi: 10.1016/j.chembiol.2020.09.003. Epub 2020 Sep 29.
Fluorescent Zn probes used for the quantitative analysis of labile Zn concentration ([Zn]) in target organelles are crucial for understanding the role of Zn in biological processes. Although several fluorescent Zn probes have been developed to date, there is still a lack of consensus concerning the [Zn] in intracellular organelles. In this study, we describe the development of ZnDA-1H, a small-molecule fluorescent probe for Zn, which exhibits less pH sensitivity, high Zn selectivity, and large fluorescence enhancement upon binding to Zn. Through protein labeling technology, ZnDA-1H was precisely targeted in various intracellular organelles, such as the nucleus, mitochondria, endoplasmic reticulum, and Golgi apparatus. ZnDA-1H exhibited a reversible fluorescence response toward labile Zn in these organelles in live cells. Using this probe, the [Zn] in the Golgi apparatus was estimated to be 25 ± 1 nM, suggesting that labile Zn plays a physiological role in the secretory pathway.
用于定量分析靶细胞器中可动锌浓度 ([Zn]) 的荧光 Zn 探针对于理解 Zn 在生物过程中的作用至关重要。尽管迄今为止已经开发了几种荧光 Zn 探针，但对于细胞内细胞器中的 [Zn] 仍然没有共识。在这项研究中，我们描述了 ZnDA-1H 的开发，这是一种用于 Zn 的小分子荧光探针，它具有较低的 pH 敏感性、高 Zn 选择性以及与 Zn 结合后荧光强度大大增强。通过蛋白质标记技术，ZnDA-1H 被精确靶向各种细胞内细胞器，如细胞核、线粒体、内质网和高尔基体。ZnDA-1H 在活细胞中这些细胞器中的可动 Zn 表现出可逆的荧光响应。使用该探针，估计高尔基体中的 [Zn] 为 25 ± 1 nM，表明可动 Zn 在分泌途径中发挥生理作用。
