Cloned (Comment) | Organism |
---|---|
gene pylS, expression as His6-tagged enzyme in Escherichia coli strain BL21 (DE3), addition of pyrrolysine to Escherichia coli cells expressing pylT, encoding tRNACUA, and pylS results in the translation of UAG in vivo as a sense codon, inability of PylS-His6 to synthesize lysyltRNACUA, co-expression with gene mtmB1 | Methanosarcina barkeri |
Protein Variants | Comment | Organism |
---|---|---|
additional information | the genetic code of Escherichia coli can be expanded to include UAG-directed pyrrolysine incorporation into proteins | Methanosarcina barkeri |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Methanosarcina barkeri |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-pyrrolysine + tRNAPyl | Methanosarcina barkeri | direct charging of tRNA(CUA) with pyrrolysine in vitro and in vivo | AMP + diphosphate + L-pyrrolysyl-tRNAPyl | - |
? | |
additional information | Methanosarcina barkeri | pyrrolysine is required in e.g. methylamine methyltransferase MtmB, overview | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Methanosarcina barkeri | - |
gene pylS | - |
Purification (Comment) | Organism |
---|---|
recombinant His6-tagged PylS from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Methanosarcina barkeri |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-pyrrolysine + tRNAPyl | direct charging of tRNA(CUA) with pyrrolysine in vitro and in vivo | Methanosarcina barkeri | AMP + diphosphate + L-pyrrolysyl-tRNAPyl | - |
? | |
ATP + L-pyrrolysine + tRNAPyl | direct charging of tRNA(CUA), isolated from Methanosarcina acetivorans strain C2A, with pyrrolysine in vitro and in vivo, PylS activates pyrrolysine with ATP and ligates pyrrolysine to tRNACUA in vitro in reactions specific for pyrrolysine | Methanosarcina barkeri | AMP + diphosphate + L-pyrrolysyl-tRNAPyl | - |
? | |
additional information | pyrrolysine is required in e.g. methylamine methyltransferase MtmB, overview | Methanosarcina barkeri | ? | - |
? | |
additional information | synthetic L-pyrrolysine is attached as a free molecule to tRNACUA by PylS, an archaeal class II aminoacyl-tRNA synthetase, inability of recombinant PylS-His6 to synthesize lysyltRNACUA | Methanosarcina barkeri | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
class II aminoacyl-tRNA synthetase | - |
Methanosarcina barkeri |
PylS | - |
Methanosarcina barkeri |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Methanosarcina barkeri |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.2 | - |
assay at | Methanosarcina barkeri |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Methanosarcina barkeri |