Cloned (Comment) | Organism |
---|---|
overexpression of the N-terminally truncated enzyme form PylRS(c270) as N-terminally His-tagged protein in Escherichia coli strain BL21(DE3), expression of selenomethionine-labeled enzyme in Escherichia coli strain B834(DE3) | Methanosarcina mazei |
Crystallization (Comment) | Organism |
---|---|
purified recombinant His-tagged N-terminally truncated enzyme form PylRS(c270) in complex with an ATP analogue AMP-PNP, hanging drop vapour diffusion method, in 100 mM sodium cacodylate, pH 6.8, containing 0.25 M NaCl, 5 mM MgSO4 and 5% w/v PEG 4000, 20°C, hexagonal crystals, X-ray diffraction structure determination and analysis at 1.9-2.6 A resolution | Methanosarcina mazei |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
33000 | - |
x * 51000, full-length enzyme, SDS-PAGE, x * 33000, recombinant N-terminally truncated enzyme form PylRS(c270), SDS-PAGE | Methanosarcina mazei |
51000 | - |
x * 51000, full-length enzyme, SDS-PAGE, x * 33000, recombinant N-terminally truncated enzyme form PylRS(c270), SDS-PAGE | Methanosarcina mazei |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-pyrrolysine + tRNAPyl | Methanosarcina mazei | - |
AMP + diphosphate + L-pyrrolysyl-tRNAPyl | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Methanosarcina mazei | - |
a methanogenic archeon, strain JCM9314 | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged N-terminally truncated enzyme form PylRS(c270) from Escherichia coli strain BL21(DE3) to homogeneity by affinity chromatography, hydrophobic interaction chromatography, and adsorption chromatography | Methanosarcina mazei |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-pyrrolysine + tRNAPyl | - |
Methanosarcina mazei | AMP + diphosphate + L-pyrrolysyl-tRNAPyl | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 51000, full-length enzyme, SDS-PAGE, x * 33000, recombinant N-terminally truncated enzyme form PylRS(c270), SDS-PAGE | Methanosarcina mazei |
More | PylRS is mainly composed of two domains: the N-terminal RNA-binding domain and the C-terminal aaRS catalytic domain | Methanosarcina mazei |
Synonyms | Comment | Organism |
---|---|---|
PylRS | - |
Methanosarcina mazei |
pyrrolysyl-tRNA synthetase | - |
Methanosarcina mazei |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Methanosarcina mazei |