2. Academic Validation
  3. 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

  • Cell Chem Biol. 2020 Dec 17;27(12):1521-1531.e8. doi: 10.1016/j.chembiol.2020.09.003.
Toshiyuki Kowada 1,  Tomomi Watanabe 2,  Yuta Amagai 3,  Rong Liu 2,  Momo Yamada 4,  Hiroto Takahashi 3,  Toshitaka Matsui 1,  Kenji Inaba 1,  Shin Mizukami 5
Affiliations

Affiliations

  • 1 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.
  • 2 Department of Molecular and Chemical Life Sciences, Graduate School of Life Sciences, Tohoku University, Sendai, Miyagi 980-8577, Japan.
  • 3 Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, Sendai, Miyagi 980-8577, Japan.
  • 4 Department of Chemistry, Graduate School of Science, Tohoku University, Sendai, Miyagi 980-8578, Japan.
  • 5 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. Electronic address: [email protected].
PMID: 32997976 DOI: 10.1016/j.chembiol.2020.09.003
Abstract

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.

Keywords

Golgi apparatus; Zn(2+); fluorescence imaging; quantitative analysis.

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