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Open AccessCommunication

Spectroscopic and Structural Analysis of Cu2+-Induced Fluorescence Quenching of ZsYellow

by In Jung Kim 1,†, Yongbin Xu 2,3 and Ki Hyun Nam 4,*
1
Division of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul 02841, Korea
2
Department of Bioengineering, College of Life Science, Dalian Minzu University, Dalian 116600, China
3
Key Laboratory of Biotechnology and Bioresources Utilization of Ministry of Education, Dalian Minzu University, Dalian 116024, China
4
Department of Life Science, Pohang University of Science and Technology, Pohang 37673, Korea
*
Author to whom correspondence should be addressed.
Present address: Department of Biotechnology and Enzyme Catalysis, Institute of Biochemistry, University of Greifswald, 17487 Greifswald, Germany.
Biosensors 2020, 10(3), 29; https://doi.org/10.3390/bios10030029
Received: 25 February 2020 / Revised: 17 March 2020 / Accepted: 19 March 2020 / Published: 23 March 2020
Fluorescent proteins exhibit fluorescence quenching by specific transition metals, suggesting their potential as fluorescent protein-based metal biosensors. Each fluorescent protein exhibits unique spectroscopic properties and mechanisms for fluorescence quenching by metals. Therefore, the metal-induced fluorescence quenching analysis of various new fluorescent proteins would be important step towards the development of such fluorescent protein-based metal biosensors. Here, we first report the spectroscopic and structural analysis of the yellow fluorescent protein ZsYellow, following its metal-induced quenching. Spectroscopic analysis showed that ZsYellow exhibited a high degree of fluorescence quenching by Cu2+. During Cu2+-induced ZsYellow quenching, fluorescence emission was recovered by adding EDTA. The crystal structure of ZsYellow soaked in Cu2+ solution was determined at a 2.6 Å resolution. The electron density map did not indicate the presence of Cu2+ around the chromophore or the β-barrel surface, which resulted in fluorescence quenching without Cu2+ binding to specific site in ZsYellow. Based on these results, we propose the fluorescence quenching to occur in a distance-dependent manner between the metal and the fluorescent protein, when these components get to a closer vicinity at higher metal concentrations. Our results provide useful insights for future development of fluorescent protein-based metal biosensors. View Full-Text
Keywords: metal biosensor; ZsYellow; fluorescent protein; fluorescence quenching; Cu2+; crystal structure metal biosensor; ZsYellow; fluorescent protein; fluorescence quenching; Cu2+; crystal structure
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Kim, I.J.; Xu, Y.; Nam, K.H. Spectroscopic and Structural Analysis of Cu2+-Induced Fluorescence Quenching of ZsYellow. Biosensors 2020, 10, 29.

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