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Literature summary for 6.1.1.26 extracted from

  • Serfling, R.; Lorenz, C.; Etzel, M.; Schicht, G.; Boettke, T.; Moerl, M.; Coin, I.
    Designer tRNAs for efficient incorporation of non-canonical amino acids by the pyrrolysine system in mammalian cells (2018), Nucleic Acids Res., 46, 1-10 .
    View publication on PubMedView publication on EuropePMC

Protein Variants

Protein Variants Comment Organism
Y349F the pyrrolysyl-tRNA synthetase/tRNAPyl pair is the most versatile and widespread system for the incorporation of non-canonical amino acids (ncAAs) into proteins in mammalian cells. Low yields of ncAA incorporation severely limit its applicability to relevant biological targets. Generation of two tRNAPyl variants that significantly boost the performance of the pyrrolysine system. Compared to the original tRNAPyl, the engineered tRNAs feature a canonical hinge between D- and T-loop, show higher intracellular concentrations and bear partially distinct post-transcriptional modifications. They allow efficient ncAA incorporation into a G-protein coupled receptor (GPCR) and simultaneous ncAA incorporation at two GPCR sites. Different combinations of recognition motifs for PylRS are introduced into Bos taurus mt-tRNASerCUA. Bicistronic plasmids are generated combining each tRNA with the PylRS from Methanosarcina barkeri, which bears the mutation Y349F for enhanced activity (PylRSF). In the presence of the PylRS substrate Lys(Boc), four tRNAs in the M series and five in the C series yield higher amber suppression compared to the tRNAPyl, whereas the other tRNAs show either lower efficiency or no amber suppression at all, overview Methanosarcina barkeri

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Methanosarcina barkeri

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + L-pyrrolysine + tRNAPyl Methanosarcina barkeri
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AMP + diphosphate + L-pyrrolysyl-tRNAPyl
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Organism

Organism UniProt Comment Textmining
Methanosarcina barkeri Q6WRH6
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-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + L-pyrrolysine + tRNAPyl
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Methanosarcina barkeri AMP + diphosphate + L-pyrrolysyl-tRNAPyl
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?
additional information the archaeal tRNAPyl features a secondary structure that significantly diverges from that of canonical tRNAs Methanosarcina barkeri ?
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?

Synonyms

Synonyms Comment Organism
PylRS
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Methanosarcina barkeri
pyrrolysyl-tRNA synthetase
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Methanosarcina barkeri

Cofactor

Cofactor Comment Organism Structure
ATP
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Methanosarcina barkeri