A Facile Platform to Engineer <i>Escherichia coli</i> Tyrosyl-tRNA Synthetase Adds New Chemistries to the Eukaryotic Genetic Code, Including a Phosphotyrosine Mimic
Katherine T. Grasso, Soumya Jyoti Singha Roy, Arianna O. Osgood, Megan J. R. Yeo, Chintan Soni, Christen M. Hillenbrand, Elise D. Ficaretta, Abhishek Chatterjee
Abstract
that overcomes this cross-reactivity. Optimized selection systems based on this tRNA enabled the efficient enrichment of both strongly and weakly active ncAA-selective EcTyrRS mutants. We also developed a wide dynamic range (WiDR) antibiotic selection to further enhance the activities of the weaker first-generation EcTyrRS mutants. We demonstrated the utility of our platform by developing several new EcTyrRS mutants that efficiently incorporated useful ncAAs in mammalian cells, including photoaffinity probes, bioconjugation handles, and a nonhydrolyzable mimic of phosphotyrosine.
Topics & Concepts
Escherichia coliMutantBioconjugationTransfer RNAGenetic codeSynthetic biologyBacterial conjugationBiochemistryBiologyComputational biologyAmino acidChemistryGeneRNARNA and protein synthesis mechanismsRNA modifications and cancerChemical Synthesis and Analysis