Minimizing the Anticodon-Recognized Loop of Methanococcus jannaschii Tyrosyl-tRNA Synthetase to Improve the Efficiency of Incorporating Noncanonical Amino Acids

Biomolecules. 2023 Mar 28;13(4):610. doi: 10.3390/biom13040610.

Abstract

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.

Keywords: anticodon recognition; genetic code expansion; giant virus; loop minimization; noncanonical amino acids.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids* / metabolism
  • Anticodon / genetics
  • Codon
  • Methanocaldococcus / genetics
  • Methanocaldococcus / metabolism
  • Tyrosine-tRNA Ligase* / genetics
  • Tyrosine-tRNA Ligase* / metabolism

Substances

  • Amino Acids
  • Tyrosine-tRNA Ligase
  • Anticodon
  • Codon

Grants and funding

This research was funded by the National Natural Science Foundation of China, grant numbers 21907107, 82073754, and 82273840 (a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions) and the National Innovation and Entrepreneurship Training Program for Undergraduate (No. 202210316060Y). The APC was funded by the National Natural Science Foundation of China, grant number 21907107.