Croatica Chemica Acta, Vol. 89 No. 2, 2016.
Izvorni znanstveni članak
https://doi.org/10.5562/cca2825
Pyrrolysyl-tRNA Synthetase, an Aminoacyl-tRNA Synthetase for Genetic Code Expansion
Ana Crnković
orcid.org/0000-0002-0581-1887
; Department of Molecular Biophysics & Biochemistry, Yale University, New Haven, CT 06520-8114, USA
Tateki Suzuki
; Department of Molecular Biophysics & Biochemistry, Yale University, New Haven, CT 06520-8114, USA
Dieter Söll
; Department of Molecular Biophysics & Biochemistry, Yale University, New Haven, CT 06520-8114, USA
Noah M. Reynolds
; Department of Molecular Biophysics & Biochemistry, Yale University, New Haven, CT 06520-8114, USA
Sažetak
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 in vitro and in vivo, they may become unique biomaterials or therapeutic agents, and they afford metabolic dependence of genetically modified organisms for biocontainment purposes. In the Methanosarcinaceae the incorporation of the 22nd genetically encoded amino acid, pyrrolysine (Pyl), is facilitated by pyrrolysyl-tRNA synthetase (PylRS) and the cognate UAG-recognizing tRNAPyl. 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 tRNAPyl 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.
This work is licensed under a Creative Commons Attribution 4.0 International License.
Ključne riječi
genetic code expansion; stop codon suppression; pyrrolysyl-tRNA synthetase; tRNAPyl; synthetic biology; non-canonical amino acid
Hrčak ID:
159878
URI
Datum izdavanja:
21.6.2016.
Posjeta: 4.336 *