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Synthesis of a highly HOCl-selective fluorescent probe and its use for imaging HOCl in cells and organisms

Title
Synthesis of a highly HOCl-selective fluorescent probe and its use for imaging HOCl in cells and organisms
Authors
Chen X.Lee K.-A.Ren X.Ryu J.-C.Kim G.Ryu J.-H.Lee W.-J.Yoon J.
Ewha Authors
윤주영Chen Xiaoqiang
SCOPUS Author ID
윤주영scopus
Issue Date
2016
Journal Title
Nature Protocols
ISSN
1754-2189JCR Link
Citation
vol. 11, no. 7, pp. 1219 - 1228
Publisher
Nature Publishing Group
Indexed
SCIE; SCOPUS WOS scopus
Abstract
During infection, nicotinamide adenine dinucleotide phosphate-oxidase of innate immune cells generates important microbicidal reactive oxygen species (ROS) such as hypochlorous acid (HOCl) to kill the invading pathogens. However, excess amounts of HOCl induce oxidative damage of functional biomolecules such as DNA and proteins, which may cause chronic inflammatory diseases. Herein, we outline protocols for the preparation of a rhodamine-based HOCl probe, as well as applications thereof, with which to detect HOCl in living cells and organisms. The probe (R19S) can be prepared from a commercially available rhodamine, rhodamine 6G, in two steps. When R19S is treated with HOCl, the sulfur atom is replaced by an oxygen atom, resulting in opening of the lactone ring; thus, nonfluorescent R19S is converted to highly fluorescent rhodamine 19 (R19). R19S exhibits high selectivity for HOCl over other ROS and high sensitivity in a weakly acidic environment. In addition, we describe fluorescence imaging assays of HOCl in mouse neutrophils and Drosophila targeted using this probe. The approximate amount of time required to synthesize the probe is 2-3 d, after which it can be used for up to 5 h in the bioimaging of living cells.
DOI
10.1038/nprot.2016.062
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자연과학대학 > 화학·나노과학전공 > Journal papers
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