Significant expansion and red-shifting of fluorescent protein chromophore determined through computational design and genetic code expansion

Li Wang; Xian Chen; Xuzhen Guo; Jiasong Li; Qi Liu; Fuying Kang; Xudong Wang; Cheng Hu; Haiping Liu; Weimin Gong; Wei Zhuang; Xiaohong Liu; Jiangyun Wang

doi:10.1007/s41048-018-0073-z

Volume 4 Issue 5

Nov. 2018

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Article Navigation > Biophysics Reports > 2018 > 4(5): 273-285

Li Wang, Xian Chen, Xuzhen Guo, Jiasong Li, Qi Liu, Fuying Kang, Xudong Wang, Cheng Hu, Haiping Liu, Weimin Gong, Wei Zhuang, Xiaohong Liu, Jiangyun Wang. Significant expansion and red-shifting of fluorescent protein chromophore determined through computational design and genetic code expansion[J]. Biophysics Reports, 2018, 4(5): 273-285. doi: 10.1007/s41048-018-0073-z

Citation:

Li Wang, Xian Chen, Xuzhen Guo, Jiasong Li, Qi Liu, Fuying Kang, Xudong Wang, Cheng Hu, Haiping Liu, Weimin Gong, Wei Zhuang, Xiaohong Liu, Jiangyun Wang. Significant expansion and red-shifting of fluorescent protein chromophore determined through computational design and genetic code expansion[J]. Biophysics Reports, 2018, 4(5): 273-285. doi: 10.1007/s41048-018-0073-z

Citation:

Li Wang, Xian Chen, Xuzhen Guo, Jiasong Li, Qi Liu, Fuying Kang, Xudong Wang, Cheng Hu, Haiping Liu, Weimin Gong, Wei Zhuang, Xiaohong Liu, Jiangyun Wang. Significant expansion and red-shifting of fluorescent protein chromophore determined through computational design and genetic code expansion[J]. Biophysics Reports, 2018, 4(5): 273-285. doi: 10.1007/s41048-018-0073-z

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Significant expansion and red-shifting of fluorescent protein chromophore determined through computational design and genetic code expansion

doi: 10.1007/s41048-018-0073-z

1 Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China
2 College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100049, China
3 Key Laboratory of Physics and Technology for Advanced Batteries(Ministry of Education), Department of Physics, Jilin University, Changchun 130012, China
4 State Key Laboratory of Structural Chemistry, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou 350002, China

Funds: This work was financially supported in part by Grants from the National Natural Science Foundation of China (21750003, 91527302, U1632133, 31628004, 21473237, and 31628004); the National Key Research and Development Program of China (2017YFA0503704 and 2016YFA0501502); and the Key Research Program of Frontier Sciences, CAS (QYZDB-SSWSMC032).

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Corresponding author: Wei Zhuang, Xiaohong Liu, Jiangyun Wang
Received Date: 24 May 2018
Publish Date: 02 November 2018

Abstract

Abstract

Fluorescent proteins (FPs) with emission wavelengths in the far-red and infrared regions of the spectrum provide powerful tools for deep-tissue and super-resolution imaging. The development of red-shifted FPs has evoked widespread interest and continuous engineering efforts. In this article, based on a computational design and genetic code expansion, we report a rational approach to significantly expand and red-shift the chromophore of green fluorescent protein (GFP). We applied computational calculations to predict the excitation and emission wavelengths of a FP chromophore harboring unnatural amino acids (UAA) and identify in silico an appropriate UAA, 2-amino-3-(6-hydroxynaphthalen-2-yl)propanoic acid (naphthol-Ala). Our methodology allowed us to formulate a GFP variant (cpsfGFP-66-Naphthol-Ala) with red-shifted absorbance and emission spectral maxima exceeding 60 and 130 nm, respectively, compared to those of GFP. The GFP chromophore is formed through autocatalytic post-translational modification to generate a planar 4-(p-hydroxybenzylidene)-5-imidazolinone chromophore. We solved the crystal structure of cpsfGFP-66-naphthol-Ala at 1.3 Å resolution and demonstrated the formation of a much larger conjugated π-system when the phenol group is replaced by naphthol. These results explain the significant red-shifting of the excitation and emission spectra of cpsfGFP-66-naphthol-Ala.
- Green fluorescent protein,
- Red-shift,
- Unnatural amino acids,
- Computational design

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