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Literature summary for 6.1.1.26 extracted from

  • Kavran, J.M.; Gundllapalli, S.; O'Donoghue, P.; Englert, M.; Soll, D.; Steitz, T.A.
    Structure of pyrrolysyl-tRNA synthetase, an archaeal enzyme for genetic code innovation (2007), Proc. Natl. Acad. Sci. USA, 104, 11268-11273.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
gene pylS, phylogenetic analysis and phylogeny of subclass IIc aaRSs, overview, expression of the N-terminally His-tagged catalytic domain of the enzyme in Escherichia coli Methanosarcina mazei

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant N-terminally His-tagged catalytic domain of PylRS complexed with either AMP-PNP, pyrrolysine-AMP plus pyrophosphate, or the pyrrolysine analogue N-epsilon-[(cylopentyloxy)carbonyl]-L-lysine plus ATP, vapour diffusion method, 10 mg/ml protein in 100 mM Tris, pH 7.0-8.0, 8-14% PEG 2000 monomethyl ether, 10 mM pyrrolysine, and10 mM AMP-PNP or other ligands, overnight at 16°C, stabilization and cryoprotection by 5 mM EDTA, 10 mM AMP-PNP, 5 mM MgCl2, 30% ethylen glycol, and additional 2% PEG, hexagonal-shaped crystals, X-ray diffraction structure determination and analysis at 1.8 A resolution Methanosarcina mazei

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+
-
Methanosarcina mazei

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + L-pyrrolysine + tRNAPyl Methanosarcina mazei pyrrolysyl-tRNA synthetase PylRS attaches L-pyrrolysine to its cognate tRNA, the special amber suppressor tRNAPyl, encoded by gene pylT AMP + diphosphate + L-pyrrolysyl-tRNAPyl
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?
additional information Methanosarcina mazei enzyme evolution study, PylRS can be placed in the aminoacyl-tRNA synthetase tree as the last known synthetase that evolved for genetic code expansion, pyrrolysine arose before the last universal common ancestral state ?
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?

Organism

Organism UniProt Comment Textmining
Methanosarcina mazei Q8PWY1 gene pylS
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Purification (Commentary)

Purification (Comment) Organism
recombinant N-terminally His-tagged catalytic domain of the enzyme from Escherichia coli by two different steps of affinity chromatography, and gel filtration Methanosarcina mazei

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + L-pyrrolysine + tRNAPyl pyrrolysyl-tRNA synthetase PylRS attaches L-pyrrolysine to its cognate tRNA, the special amber suppressor tRNAPyl, encoded by gene pylT Methanosarcina mazei AMP + diphosphate + L-pyrrolysyl-tRNAPyl
-
?
ATP + L-pyrrolysine + tRNAPyl substrate binding, PylRS utilizes a deep hydrophobic pocket for recognition of the Pyl side chain Methanosarcina mazei AMP + diphosphate + L-pyrrolysyl-tRNAPyl pyrrolysine-AMPbinds in a deep hydrophobic pocket, with its position coordinated by a hydrogen-bonding network with PylRS, binding structure, overview ?
additional information enzyme evolution study, PylRS can be placed in the aminoacyl-tRNA synthetase tree as the last known synthetase that evolved for genetic code expansion, pyrrolysine arose before the last universal common ancestral state Methanosarcina mazei ?
-
?
additional information substrate-binding specificity of PylRS, overview Methanosarcina mazei ?
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?

Synonyms

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

Cofactor

Cofactor Comment Organism Structure
ATP
-
Methanosarcina mazei