An aminoacyl-tRNA synthetase that specifically activates pyrrolysine.

@article{Polycarpo2004AnAS,
  title={An aminoacyl-tRNA synthetase that specifically activates pyrrolysine.},
  author={C. Polycarpo and A. Ambrogelly and A. B{\'e}rub{\'e} and SusAnn M. Winbush and J. McCloskey and P. Crain and J. L. Wood and D. S{\"o}ll},
  journal={Proceedings of the National Academy of Sciences of the United States of America},
  year={2004},
  volume={101 34},
  pages={
          12450-4
        }
}
Pyrrolysine, the 22nd cotranslationally inserted amino acid, was found in the Methanosarcina barkeri monomethylamine methyltransferase protein in a position that is encoded by an in-frame UAG stop codon in the mRNA. M. barkeri encodes a special amber suppressor tRNA (tRNA(Pyl)) that presumably recognizes this UAG codon. It was reported that Lys-tRNA(Pyl) can be formed by the aminoacyl-tRNA synthetase-like M. barkeri protein PylS [Srinivasan, G., James, C. M. & Krzycki, J. A. (2002) Science 296… Expand
Recognition of non-alpha-amino substrates by pyrrolysyl-tRNA synthetase.
TLDR
It is reported here that wild type PylRS can recognize substrates with a variety of main-chain alpha-groups: alpha-hydroxyacid, non-alpha-amino-carboxylic acid, N(alpha)-methyl-amINO acid, and D-aminos acid, each with the same side chain as that of Boc-lysine, in contrast, PheRS recognizes none of these amino acid analogs. Expand
Pyrrolysyl-tRNA synthetase:tRNAPyl structure reveals the molecular basis of orthogonality
TLDR
It is shown that Desulfitobacterium hafniense PylRS–tRNAPyl is an orthogonal pair in vitro and in vivo, and the crystal structure of this orthogonality pair is presented. Expand
Pyrrolysine is not hardwired for cotranslational insertion at UAG codons
TLDR
Data suggest that t RNAPyl variants may decode numerous codons and that tRNAPyl:PylRS is a fine orthogonal tRNA:synthetase pair that facilitated the late addition of Pyl to the genetic code. Expand
Recognition of pyrrolysine tRNA by the Desulfitobacterium hafniense pyrrolysyl-tRNA synthetase
TLDR
The tRNAPyl structure contains the highly conserved T-loop contact U54·A58 and position 57 is conserved as a purine, but the canonical T- to D- loop contact between positions 18 and 56 was not present and the tRN APyl anticodon was shown not to be important for recognition by bacterial PylRS. Expand
Specificity of pyrrolysyl-tRNA synthetase for pyrrolysine and pyrrolysine analogs.
TLDR
Interactions necessary for recognition of substrates by archaeal PylS are probed via synthesis of close pyrrolysine analogs and testing their reactivity in amino acid activation assays, illustrating the relative importance of the H-bonding and hydrophobic interactions in the recognition of the methylpyrroline ring of pyr rolysine. Expand
The appearance of pyrrolysine in tRNAHis guanylyltransferase by neutral evolution
TLDR
Phylogenetic analysis suggests that Thg1 is becoming dispensable in the archaea, and supports the hypothesis that Pyl appeared in MaThg1 as the result of neutral evolution, indicating that even the most unusual amino acid can play an ordinary role in proteins. Expand
Misacylation of pyrrolysine tRNA in vitro and in vivo
TLDR
It is shown that M. barkeri Fusaro tRNAPyl can be misacylated with serine by the M. barkingeri bacterial‐type seryl‐tRNA synthetase in vitro and in vivo in Escherichia coli. Expand
Translation of UAG as Pyrrolysine
TLDR
Pyrrolysine incorporation appears to occur to some extent by amber suppression on a genome-wide basis in methanogenic Archaea, and some methanogen genomes encode additional homologs of elongation and release factors, however, their limited distribution suggests at best a nonessential role in enhancing UAG translation as pyrrolesine. Expand
The complete biosynthesis of the genetically encoded amino acid pyrrolysine from lysine
TLDR
The results indicate that the radical S-adenosyl-l-methionine (SAM) protein PylB mediates a lysine mutase reaction that produces 3-methylornithine, which is then ligated to a second molecule of l Lysine by PylC before oxidation byPylD results in pyrrolysine. Expand
Structure of pyrrolysyl-tRNA synthetase, an archaeal enzyme for genetic code innovation
TLDR
Three crystal structures of the Methanosarcina mazei PylRS complexed with either AMP–PNP, Pyl–AMP plus pyrophosphate, or the Pyl analogue N-ε-[(cylopentyloxy)carbonyl]-l-lysine plus ATP are determined, revealing that Pyrrolysine becomes attached to its cognate tRNA by pyrrolysyl-tRNA synthetase (PylRS). Expand
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References

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TLDR
Results indicate that pyrrolysine is the 22nd genetically encoded natural amino acid to be encoded by the UAG codon in methylamine methyltransferase genes of Methanosarcina barkeri. Expand
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TLDR
Mutation S294 was the key for proposing a mechanism by which the substrate analogue p-F-Phe is excluded from the enzymatic reaction; this may be achieved by steric interactions between the para-position of the aromatic ring and the amino acid residue at position 294. Expand
Activation of the pyrrolysine suppressor tRNA requires formation of a ternary complex with class I and class II lysyl-tRNA synthetases.
TLDR
It is shown that tRNAPyl is efficiently aminoacylated in the presence of both the class I LysRS and class II LysRS of M. barkeri, but not by either enzyme acting alone or by PylS. Expand
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TLDR
Analysis of tRNA isolated under acidic conditions showed that this novel modification is present in normal E. coli tRNA; presumably it previously escaped detection as the standard conditions of t RNA isolation include an alkaline deacylation step that also causes hydrolysis of glutamyl-queuosine. Expand
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TLDR
Comparison of the pure enzymes LysS and LysU indicates that, in the presence of saturating substrates, LysS is about twice more active than LysU in the ATP-PPi exchange as well as in the tRNA aminoacylation reaction, and this observation points to a possible useful role of LysU, under physiological conditions causing cadaverine accumulation in the bacterium. Expand
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TLDR
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TLDR
The UAG-encoded residue in a 1.55 angstrom resolution structure of the Methanosarcina barkerimonomethylamine methyltransferase (MtmB) reveals a homohexamer comprised of individual subunits with a TIM barrel fold that appears consistent with a lysine in amide-linkage to (4R,5R)-4-substituted-pyrroline-5-carboxylate. Expand
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TLDR
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TLDR
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