Label-Free Fluorometric Method for Monitoring Conformational Flexibility of Laccase Based on a Selective Laccase Sensor
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- 作者：Suyan Qiu ; Zhenyu Lin ; Yaomin Zhou ; Ruili Li ; Jinyan Zhang ; Dawen Zhang ; Linguang Luo ; Longhua Guo ; Bin Qiu ; Guonan Chen
- 刊名：Analytical Chemistry
- 出版年：2013
- 出版时间：November 19, 2013
- 年：2013
- 卷：85
- 期：22
- 页码：11041-11046
- 全文大小：364K
- 年卷期：v.85,no.22(November 19, 2013)
- ISSN：1520-6882
文摘
A facile and selective fluorescence sensor for laccase determination has been proposed depending on the interaction between 3-azidocoumarin and trametes versicolor (Tv) laccase in this paper. The azido group of 3-azidocoumarin that is electron-rich 伪-nitrogen can directly interact with histidines that coordinate to three copper sites through hydrogen bonds and forms a new complex, which decreases the electron-donating ability of the azido group, leading to enhance the fluorescence intensity of the sensing system. Also, other common proteins have no significant interference for the proposed laccase sensor. Additionally, the proposed fluorescence sensor is extended to demonstrate the conformational flexibility of Tv laccase by the urea denaturant. A good consistency of the results obtained with the presented laccase sensor and CD spectra is performed. Furthermore, the relationship between the catalytic activity and the unfolding percentage of the unfolded Tv laccase through the proposed laccase sensor is also elucidated well.