Pyrrolysyl-tRNA synthetase, an aminoacyl-tRNA synthetase for genetic code expansion. [PDF]
Croat Chem Acta, 2016 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.
Crnković A +3 more
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Ferritin Conjugates With Multiple Clickable Amino Acids Encoded by C-Terminal Engineered Pyrrolysyl-tRNA Synthetase. [PDF]
Front Chem, 2021 This study reports the application of expanding genetic codes in developing protein cage-based delivery systems. The evolved Methanosarcina mazei pyrrolysyl-tRNA synthetase (PylRS)•tRNAPyl pairs derived from directed evolution are examined to probe their recognition for para-substituted phenylalanine analogs.
Wang YH +5 more
europepmc +5 more sources
Pyrrolysyl-tRNA synthetase variants reveal ancestral aminoacylation function. [PDF]
FEBS Lett, 2013 Pyrrolysyl‐tRNA synthetase (PylRS) is a class IIc aminoacyl‐tRNA synthetase that is related to phenylalanyl‐tRNA synthetase (PheRS). Genetic selection provided PylRS variants with a broad range of specificity for diverse non‐canonical amino acids (ncAAs). One variant is a specific phenylalanine‐incorporating enzyme.
Ko JH +5 more
europepmc +4 more sources
Engineering a Polyspecific Pyrrolysyl-tRNA Synthetase by a High Throughput FACS Screen. [PDF]
Sci Rep, 2019 AbstractThe Pyrrolysyl-tRNA synthetase (PylRS) and its cognate tRNAPyl are extensively used to add non-canonical amino acids (ncAAs) to the genetic code of bacterial and eukaryotic cells. However, new ncAAs often require a cumbersome de novo engineering process to generate an appropriate PylRS/tRNAPyl pair.
Hohl A +8 more
europepmc +6 more sources
Update of the Pyrrolysyl-tRNA Synthetase/tRNAPyl Pair and Derivatives for Genetic Code Expansion. [PDF]
J Bacteriol, 2023 The cotranslational incorporation of pyrrolysine (Pyl), the 22nd proteinogenic amino acid, into proteins in response to the UAG stop codon represents an outstanding example of natural genetic code expansion. Genetic encoding of Pyl is conducted by the pyrrolysyl-tRNA synthetase (PylRS) and its cognate tRNA, tRNA Pyl
Gong X, Zhang H, Shen Y, Fu X.
europepmc +3 more sources
Crystal structures reveal an elusive functional domain of pyrrolysyl-tRNA synthetase. [PDF]
Nat Chem Biol, 2017 Pyrrolysyl-tRNA synthetase (PylRS) is a major tool in genetic code expansion using noncanonical amino acids, yet its structure and function are not completely understood. Here we describe the crystal structure of the previously uncharacterized essential N-terminal domain of this unique enzyme in complex with tRNAPyl.
Suzuki T +7 more
europepmc +5 more sources
A Designed, Highly Efficient Pyrrolysyl-tRNA Synthetase Mutant Binds o-Chlorophenylalanine Using Two Halogen Bonds. [PDF]
J Mol Biol, 2022 As one of the most valuable tools for genetic code expansion, pyrrolysyl-tRNA synthetase (PylRS) is structurally related to phenylalanyl-tRNA synthetase (PheRS). By introducing mutations that mimic ligand interactions in PheRS into PylRS, we designed a PylRS mutant.
Vatansever EC +6 more
europepmc +3 more sources
Transferability of N-terminal mutations of pyrrolysyl-tRNA synthetase in one species to that in another species on unnatural amino acid incorporation efficiency. [PDF]
Amino Acids, 2021 Genetic code expansion is a powerful technique for site-specific incorporation of an unnatural amino acid into a protein of interest. This technique relies on an orthogonal aminoacyl-tRNA synthetase/tRNA pair and has enabled incorporation of over 100 ...
Williams TL +5 more
europepmc +4 more sources
Chimeric design of pyrrolysyl-tRNA synthetase/tRNA pairs and canonical synthetase/tRNA pairs for genetic code expansion. [PDF]
Nat Commun, 2020 AbstractAn orthogonal aminoacyl-tRNA synthetase/tRNA pair is a crucial prerequisite for site-specific incorporation of unnatural amino acids. Due to its high codon suppression efficiency and full orthogonality, the pyrrolysyl-tRNA synthetase/pyrrolysyl-tRNA pair is currently the ideal system for genetic code expansion in both eukaryotes and prokaryotes.
Ding W +7 more
europepmc +5 more sources
Genetic incorporation of histidine derivatives using an engineered pyrrolysyl-tRNA synthetase. [PDF]
ACS Chem Biol, 2014 A polyspecific amber suppressor aminoacyl-tRNA synthetase/tRNA pair was evolved that genetically encodes a series of histidine analogues in both Escherichia coli and mammalian cells. In combination with tRNACUA(Pyl), a pyrrolysyl-tRNA synthetase mutant was able to site-specifically incorporate 3-methyl-histidine, 3-pyridyl-alanine, 2-furyl-alanine, and
Xiao H +5 more
europepmc +3 more sources