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 (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 | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Methanosarcina mazei |
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 | - |
? | |
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 | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Methanosarcina mazei | Q8PWY1 | gene pylS | - |
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 | 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 | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
PylRS | - |
Methanosarcina mazei |
pyrrolysyl-tRNA synthetase | - |
Methanosarcina mazei |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Methanosarcina mazei |