Evolving the N-Terminal Domain of Pyrrolysyl-tRNA Synthetase for Improved Incorporation of Noncanonical Amino Acids. Academic Article uri icon

abstract

  • By evolving the N-terminal domain of Methanosarcina mazei pyrrolysyl-tRNA synthetase (PylRS) that directly interacts with tRNAPyl , a mutant clone displaying improved amber-suppression efficiency for the genetic incorporation of N -(tert-butoxycarbonyl)-l-lysine threefold more than the wild type was identified. The identified mutations were R19H/H29R/T122S. Direct transfer of these mutations to two other PylRS mutants that were previously evolved for the genetic incorporation of N -acetyl-l-lysine and N -(4-azidobenzoxycarbonyl)-l-,-dehydrolysine also improved the incorporation efficiency of these two noncanonical amino acids. As the three identified mutations were found in the N-terminal domain of PylRS that was separated from its catalytic domain for charging tRNAPyl with a noncanonical amino acid, they could potentially be introduced to all other PylRS mutants to improve the incorporation efficiency of their corresponding noncanonical amino acids. Therefore, it represents a general strategy to optimize the pyrrolysine incorporation system-based noncanonical amino-acid mutagenesis.

published proceedings

  • Chembiochem

altmetric score

  • 1.75

author list (cited authors)

  • Sharma, V., Zeng, Y. u., Wang, W. W., Qiao, Y., Kurra, Y., & Liu, W. R.

citation count

  • 13

complete list of authors

  • Sharma, Vangmayee||Zeng, Yu||Wang, W Wesley||Qiao, Yuchen||Kurra, Yadagiri||Liu, Wenshe R

publication date

  • January 2018

publisher