Quantitative Imaging of Labile Zn2+ in the Golgi Apparatus Using a Localizable Small-Molecule Fluorescent Probe
Quantitative Imaging of Labile Zn2+ in the Golgi Apparatus Using a Localizable Small-Molecule Fluorescent Probe
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DOI：
10.1016/j.chembiol.2020.09.003
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发表时间：
2020-12-17
影响因子：
8.6
通讯作者：
Mizukami, Shin
中科院分区：
文献类型：
--
作者：
Kowada, Toshiyuki;Watanabe, Tomomi;Mizukami, Shin
用于定量分析靶细胞器中不稳定的锌离子浓度的荧光锌离子探针对于理解锌离子在生物过程中的作用是至关重要的。尽管到目前为止已经开发了几种锌离子荧光探针，但对于细胞内细胞器中的[锌离子]仍然缺乏共识。在本研究中，我们介绍了一种小分子锌离子荧光探针--ZnDA-1H的研制，它具有较低的pH灵敏度、较高的锌离子选择性和与锌离子结合后较大的荧光增强作用。通过蛋白质标记技术，可以精确定位于细胞核、线粒体、内质网和高尔基体等多种细胞内细胞器。在活细胞中，对这些细胞器中的不稳定的锌离子表现出可逆的荧光响应。用该探针测得高尔基体的[锌离子]为25+/-1 nm，表明不稳定的锌离子在分泌途径中起着生理作用。
Fluorescent Zn2+ probes used for the quantitative analysis of labile Zn2+ concentration ([Zn2+]) in target organelles are crucial for understanding the role of Zn2+ in biological processes. Although several fluorescent Zn2+ probes have been developed to date, there is still a lack of consensus concerning the [Zn2+] in intracellular organelles. In this study, we describe the development of ZnDA-1H, a small-molecule fluorescent probe for Zn2+, which exhibits less pH sensitivity, high Zn2+ selectivity, and large fluorescence enhancement upon binding to Zn2+. 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 Zn2+ in these organelles in live cells. Using this probe, the [Zn2+] in the Golgi apparatus was estimated to be 25 +/- 1 nM, suggesting that labile Zn2+ plays a physiological role in the secretory pathway.