Quantitative Imaging of Labile Zn2+ in the Golgi Apparatus Using a Localizable Small-Molecule Fluorescent Probe
| Autor: | Yuta Amagai, Hiroto Takahashi, Kenji Inaba, Shin Mizukami, Toshitaka Matsui, Tomomi Watanabe, Momo Yamada, Rong Liu, Toshiyuki Kowada |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
inorganic chemicals
Fluorescence-lifetime imaging microscopy Clinical Biochemistry Biology 01 natural sciences Biochemistry symbols.namesake Drug Discovery Organelle medicine Molecular Biology Secretory pathway Pharmacology 010405 organic chemistry Endoplasmic reticulum Golgi apparatus Small molecule Fluorescence 0104 chemical sciences enzymes and coenzymes (carbohydrates) medicine.anatomical_structure biological sciences health occupations symbols Biophysics bacteria Molecular Medicine Nucleus |
| Zdroj: | Cell Chemical Biology. 27:1521-1531.e8 |
| ISSN: | 2451-9456 |
| DOI: | 10.1016/j.chembiol.2020.09.003 |
| Popis: | Summary 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. |
| Databáze: | OpenAIRE |
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