Aminoacyl-tRNA synthetases are crucial for maintaining the fidelity of the translation of the genetic code by ensuring specificity for their cognate substrates and are remarkable in that they constitute a set of proteins capable of binding each of the 20 canonical amino acids. As such, they represent an important class of proteins that may be of great use for a myriad of bioengineering applications. Indeed, recent advances in re-engineering tyrosyl-tRNA synthtease has allowed for the incorporation of non-natural amino acids into proteins. However, while the anticodon binding domains have been well studied, comparatively little is known about the specificity and binding affinity of the conserved amino acid binding domains for each of the aa-tRNA synthetases in isolation (i.e. separate from downstream effects on the aminoacylation activity). In this work we characterize the wild-type binding affinity of the conserved catalytic core region of several tRNA synthetases for different substrates. We also demonstrate the use of directed evolution using semi-rational design and implementation of advanced library generation with phage display screening techniques for the use in evolving improved amino acid biosensors.

Characterization and directed evolution of aminoacyl tRNA synthetases

249th National Meeting (2015)

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