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Pyrrolysyl-tRNA Synthetase, an Aminoacyl-tRNA Synthetase for Genetic Code Expansion

Journal Article · · Croatica Chemica Acta
DOI:https://doi.org/10.5562/cca2825· OSTI ID:1434637
 [1];  [2];  [2];  [2]
  1. Yale Univ., New Haven, CT (United States); None
  2. Yale Univ., New Haven, CT (United States)
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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.
Research Organization:
Michigan State Univ., East Lansing, MI (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
Grant/Contract Number:
FG02-98ER20311
OSTI ID:
1434637
Journal Information:
Croatica Chemica Acta, Journal Name: Croatica Chemica Acta Journal Issue: 2 Vol. 89; ISSN 0011-1643
Publisher:
The Croatian Chemical SocietyCopyright Statement
Country of Publication:
United States
Language:
English

Cited By (7)

Crystal structures reveal an elusive functional domain of pyrrolysyl-tRNA synthetase journal October 2017
Auxotrophy to Xeno-DNA: an exploration of combinatorial mechanisms for a high-fidelity biosafety system for synthetic biology applications collection January 2018
Recent Development of Genetic Code Expansion for Posttranslational Modification Studies journal July 2018
Pyrrolysyl-tRNA Synthetase with a Unique Architecture Enhances the Availability of Lysine Derivatives in Synthetic Genetic Codes journal September 2018
tRNA Pyl : Structure, function, and applications journal September 2017
Understanding the Genetic Code journal April 2019
Auxotrophy to Xeno-DNA: an exploration of combinatorial mechanisms for a high-fidelity biosafety system for synthetic biology applications journal August 2018

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Related Subjects

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY