Pyrrolysyl-tRNA Synthetase, an Aminoacyl-tRNA Synthetase for Genetic Code Expansion
Genetic code expansion (GCE) has become a central topic of synthetic biology. GCE relies on engineered aminoacyl‐tRNA synthetases (aaRSs) and a cognate tRNA species to allow codon reassignment by co-translational insertion of non-canonical amino acids (ncAAs) into proteins. Introduction of such amin...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Croatian Chemical Society
2016-06-01
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| Series: | Croatica Chemica Acta |
| Online Access: | http://hrcak.srce.hr/file/235773 |
| Summary: | Genetic code expansion (GCE) has become a central topic of synthetic biology. GCE relies on engineered aminoacyl‐tRNA synthetases (aaRSs) and a cognate tRNA species to allow codon reassignment by co-translational insertion of non-canonical amino acids (ncAAs) into proteins. Introduction of such amino acids increases the chemical diversity of recombinant proteins endowing them with novel properties. Such proteins serve in sophisticated biochemical and biophysical studies both <i>in vitro</i> and <i>in vivo</i>, they may become unique biomaterials or therapeutic agents, and they afford metabolic dependence of genetically modified organisms for biocontainment purposes. In the <i>Methanosarcinaceae</i> the incorporation of the 22<sup>nd</sup> genetically encoded amino acid, pyrrolysine (Pyl), is facilitated by pyrrolysyl-tRNA synthetase (PylRS) and the cognate UAG-recognizing tRNA<sup>Pyl</sup>. This unique aaRS⋅tRNA pair functions as an orthogonal translation system (OTS) in most model organisms. The facile directed evolution of the large PylRS active site to accommodate many ncAAs, and the enzyme’s anticodon-blind specific recognition of the cognate tRNA<sup>Pyl</sup> make this system highly amenable for GCE purposes. The remarkable polyspecificity of PylRS has been exploited to incorporate >100 different ncAAs into proteins. Here we review the Pyl-OT system and selected GCE applications to examine the properties of an effective OTS.
<br><a rel="license" href="http://creativecommons.org/licenses/by/4.0/"><img alt="Creative Commons License" style="border-width:0" src="https://i.creativecommons.org/l/by/4.0/80x15.png" /></a> This work is licensed under a <a rel="license" href="http://creativecommons.org/licenses/by/4.0/">Creative Commons Attribution 4.0 International License</a>. |
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| ISSN: | 0011-1643 1334-417X |