Quantitative Imaging of Labile Zn2+ in the Golgi Apparatus Using a Localizable Small-Molecule Fluorescent Probe.

Toshiyuki Kowada, Tomomi Watanabe, Yuta Amagai, Rong Liu, Momo Yamada, Hiroto Takahashi, Toshitaka Matsui, Kenji Inaba, Shin Mizukami

Cell chemical biology 2020 Dec 17

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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. Copyright © 2020 Elsevier Ltd. All rights reserved.

Citation

Toshiyuki Kowada, Tomomi Watanabe, Yuta Amagai, Rong Liu, Momo Yamada, Hiroto Takahashi, Toshitaka Matsui, Kenji Inaba, Shin Mizukami. Quantitative Imaging of Labile Zn2+ in the Golgi Apparatus Using a Localizable Small-Molecule Fluorescent Probe. Cell chemical biology. 2020 Dec 17;27(12):1521-1531.e8