Minimizing the Anticodon-Recognized Loop of <i>Methanococcus jannaschii</i> Tyrosyl-tRNA Synthetase to Improve the Efficiency of Incorporating Noncanonical Amino Acids
Zhiyang Hu,
Jinming Liang,
Taogeng Su,
Di Zhang,
Hao Li,
Xiangdong Gao,
Wenbin Yao,
Xiaoda Song
Affiliations
Zhiyang Hu
Jiangsu Key Laboratory of Druggability of Biopharmaceuticals, Department of Biochemistry, School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, China
Jinming Liang
Jiangsu Key Laboratory of Druggability of Biopharmaceuticals, Department of Biochemistry, School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, China
Taogeng Su
Jiangsu Key Laboratory of Druggability of Biopharmaceuticals, Department of Biochemistry, School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, China
Di Zhang
Jiangsu Key Laboratory of Druggability of Biopharmaceuticals, Department of Biochemistry, School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, China
Hao Li
Jiangsu Key Laboratory of Druggability of Biopharmaceuticals, Department of Biochemistry, School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, China
Xiangdong Gao
Jiangsu Key Laboratory of Druggability of Biopharmaceuticals, Department of Biochemistry, School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, China
Wenbin Yao
Jiangsu Key Laboratory of Druggability of Biopharmaceuticals, Department of Biochemistry, School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, China
Xiaoda Song
Jiangsu Key Laboratory of Druggability of Biopharmaceuticals, Department of Biochemistry, School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, China
In the field of genetic code expansion (GCE), improvements in the efficiency of noncanonical amino acid (ncAA) incorporation have received continuous attention. By analyzing the reported gene sequences of giant virus species, we noticed some sequence differences at the tRNA binding interface. On the basis of the structural and activity differences between Methanococcus jannaschii Tyrosyl-tRNA Synthetase (MjTyrRS) and mimivirus Tyrosyl-tRNA Synthetase (MVTyrRS), we found that the size of the anticodon-recognized loop of MjTyrRS influences its suppression activity regarding triplet and specific quadruplet codons. Therefore, three MjTyrRS mutants with loop minimization were designed. The suppression of wild-type MjTyrRS loop-minimized mutants increased by 1.8–4.3-fold, and the MjTyrRS variants enhanced the activity of the incorporation of ncAAs by 15–150% through loop minimization. In addition, for specific quadruplet codons, the loop minimization of MjTyrRS also improves the suppression efficiency. These results suggest that loop minimization of MjTyrRS may provide a general strategy for the efficient synthesis of ncAAs-containing proteins.
