Developing a genetically encoded green fluorescent protein mutant for sensitive light-up fluorescent sensing and cellular imaging of Hg(II)

被引:21
|
作者
Jiang, Tao [1 ]
Guo, Daiping [1 ]
Wang, Qian [1 ]
Wu, Xin [1 ]
Li, Zhao [1 ]
Zheng, Zhenhua [2 ]
Yin, Boyuan [2 ]
Xia, Lin [2 ]
Tang, Jixian [2 ]
Luo, Wenxin [2 ]
Xia, Ningshao [2 ]
Jiang, Yunbao [1 ]
机构
[1] Xiamen Univ, Dept Chem, Coll Chem & Chem Engn, MOE Key Lab Analyt Sci, Xiamen 361005, Peoples R China
[2] Xiamen Univ, State Key Lab Mol Vaccinol & Mol Diagnost, Natl Inst Diagnost & Vaccine Dev Infect Dis, Sch Life Sci, Xiamen 361005, Peoples R China
关键词
Green fluorescent protein-based sensor; Mercury ion; Fluorescence light-up; Cellular imaging; STRUCTURAL BASIS; DYNAMICS; VARIANTS; MERCURY; HG2+; DESIGN; SENSOR; ION; RECOGNITION; EXCITATION;
D O I
10.1016/j.aca.2015.03.026
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Hg(II) is well-known for quenching fluorescence in a distance dependent manner. Nevertheless, when we exposed the fluorophore of a green fluorescent protein (GFP) toward Hg(II), through H148C mutation, the GFP fluorescence could be "lighted up" by Hg(II) down to sub-nM level. The detection linear range is 0.5-3.0 nM for protein solutions at 8.0 nM. The GFPH148C protein displayed a promising selectivity toward Hg(II) and also the cellular imaging capacity. Spectra measurements suggested that the ground-state redistribution of protein contributed to the fluorescence enhancement, which was found not limited to Hg(II), and thus presented an opening for building a pool of GFP-based chemosensors toward other heavy metal ions. (C) 2015 Elsevier B.V. All rights reserved.
引用
收藏
页码:77 / 82
页数:6
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