Quantitative Imaging of Labile Zn2+ in the Golgi Apparatus Using a Localizable Small-Molecule Fluorescent Probe. Issue 12 (17th December 2020)

• Record Type: Journal Article
• Authors: Kowada, Toshiyuki; Watanabe, Tomomi; Amagai, Yuta; Liu, Rong; Yamada, Momo; Takahashi, Hiroto; Matsui, Toshitaka; Inaba, Kenji; Mizukami, Shin
• Is Part Of: Cell chemical biology. Volume 27:Issue 12(2020)
• Page Start: 1521
• Page End: 1531.e8
• Publication Date: 2020-12-17
• Subjects: Zn2+ -- quantitative analysis -- fluorescence imaging -- Golgi apparatus; Biochemistry -- Periodicals; 572.05
• DOI: 10.1016/j.chembiol.2020.09.003
• Languages: English
• ISSNs: 2451-9456

Abstract

Summary: Fluorescent Zn 2+ probes used for the quantitative analysis of labile Zn 2+ concentration ([Zn 2+ ]) in target organelles are crucial for understanding the role of Zn 2+ in biological processes. Although several fluorescent Zn 2+ probes have been developed to date, there is still a lack of consensus concerning the [Zn 2+ ] in intracellular organelles. In this study, we describe the development of ZnDA-1H, a small-molecule fluorescent probe for Zn 2+, which exhibits less pH sensitivity, high Zn 2+ selectivity, and large fluorescence enhancement upon binding to Zn 2+ . 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 2+ in these organelles in live cells. Using this probe, the [Zn 2+ ] in the Golgi apparatus was estimated to be 25 ± 1 nM, suggesting that labile Zn 2+ plays a physiological role in the secretory pathway. Graphical Abstract: Highlights: ZnDA-1H is a "turn-on" type green fluorescent probe with high Zn 2+ selectivity The K d toward Zn 2+ and emission of ZnDA-1H are less dependent on pH change ZnDA-1H specifically localizes to target organelles upon conjugating with HaloTag ZnDA-1H was applied to quantify the [Zn 2+ ] in the Golgi apparatus Abstract : Kowada et al. have developed a small-molecule fluorescent Zn 2+ probe, ZnDA-1H, which can be localized specifically to intracellular organelles by utilizing a protein labeling technology. ZnDA-1H enables quantitative analysis of labile Zn 2+ in the Golgi apparatus.