The characteristic study of peroxynitrite fluorescent probe Ds-DAB

• Main Authors: Lin, Ku-Kuei, 林谷奎
• Other Authors: Wang, Yun-Ming
• Format: Others
• Language: zh-TW
• Published: 2013
• Online Access: http://ndltd.ncl.edu.tw/handle/44053117391022011250

碩士 === 國立交通大學 === 生物科技系所 === 101 === Peroxynitrite (ONOO－) is an important signaling molecule in organism. It is involved in diverse physiological and pathological pathways. Visualization of peroxynitrite using bioimaging techniques provide us to advance the understanding of roles played by peroxynitrite in organism. The fluorescent ONOO－probe, Ds-DAB, was developed in this study. The design strategy of Ds-DAB is based on the peroxynitrite anion might react with secondary amine to form nitrosamine by direct nucleophilic nitrosation. Then, the benzotriazol was released and formed dansyl acid with high fluorescence intensity. Ds-DAB has not only fast reaction rate (20 sec.) but high specificity for peroxynitrite over other reactive nitrogen species and oxygen species. Besides, an excellent linearity between the fluorescence intensity and peroxynitrite concentration could provide the quantitative measurement through fluorometric method. The detection limit was 52.4 nM. In addition, Ds-DAB can be applied to monitor the endogenous peroxynitrite with low cytotoxicity. Ds-DAB had hardly any effect on the fluorescence intensity over the pH range from 5.8 to 11.9. Therefore, the probe is expected to work well under physiological condition.