Significant improvement of oxidase activity through the genetic incorporation of a redox-active unnatural amino acid
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| Authors: | Yang Yu Qing Zhou Li Wang Xiaohong Liu Wei Zhang Meirong Hu Jianshu Dong Jiasong Li Xiaoxuan Lv Hanlin Ouyang Han Li Feng Gao Weimin Gong Yi Lu Jiangyun Wang |
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| Affiliation: | a School of Life Sciences , University of Science and Technology of China , Hefei , Anhui 230026 , China ; b Laboratory of RNA Biology , Institute of Biophysics , Chinese Academy of Sciences , 15 Datun Road, Chaoyang District , Beijing , 100101 , China . Email: ; c Center of Biophysics and Computational Biology and Department of Chemistry , University of Illinois at Urbana-Champaign , Urbana , Illinois 61801 , USA . Email: |
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| Abstract: | While nature employs various covalent and non-covalent strategies to modulate tyrosine (Y) redox potential and pKa in order to optimize enzyme activities, such approaches have not been systematically applied for the design of functional metalloproteins. Through the genetic incorporation of 3-methoxytyrosine (OMeY) into myoglobin, we replicated important features of cytochrome c oxidase (CcO) in this small soluble protein, which exhibits selective O2 reduction activity while generating a small amount of reactive oxygen species (ROS). These results demonstrate that the electron donating ability of a tyrosine residue in the active site is important for CcO function. Moreover, we elucidated the structural basis for the genetic incorporation of OMeY into proteins by solving the X-ray structure of OMeY specific aminoacyl-tRNA synthetase complexed with OMeY. |
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