A Designed, Highly Efficient Pyrrolysyl-tRNA Synthetase Mutant Binds o-Chlorophenylalanine Using Two Halogen Bonds. Academic Article uri icon


  • As one of the most valuable tools for genetic code expansion, pyrrolysyl-tRNA synthetase (PylRS) is structurally related to phenylalanyl-tRNA synthetase (PheRS). By introducing mutations that mimic ligand interactions in PheRS into PylRS, we designed a PylRS mutant. This mutant, designated as oClFRS, recognizes a number of o-substituted phenylalanines for their genetic incorporation at amber codon. Its efficiency in catalyzing genetic incorporation of o-chlorophenylalanine (o-ClF) is better than that for N-tert-butyloxycarbonyl-lysine catalyzed by PylRS. The crystal structure of oClFRS bound with o-ClF shows that o-ClF binds deeply into a hydrophobic but catalytically inactive pocket in the active site and involves two halogen bonds to achieve strong interactions. The shift of o-ClF to a catalytically active position in the oClFRS active site will be necessary for its activation. This is the first reported aminoacyl-tRNA synthetase that involves two halogen bonds for ligation recognition and might represent an alternative route to develop aminoacyl-tRNA synthetase mutants that are selective for noncanonical amino acids over native amino acids.

published proceedings

  • J Mol Biol

altmetric score

  • 5.1

author list (cited authors)

  • Vatansever, E. C., Yang, K. S., Geng, Z. Z., Qiao, Y., Li, P., Xu, S., & Liu, W. R.

citation count

  • 4

complete list of authors

  • Vatansever, Erol C||Yang, Kai S||Geng, Zhi Zachary||Qiao, Yuchen||Li, Pingwei||Xu, Shiqing||Liu, Wenshe Ray

publication date

  • April 2022