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Literature summary for 6.1.1.26 extracted from

  • Blight, S.K.; Larue, R.C.; Mahapatra, A.; Longstaff, D.G.; Chang, E.; Zhao, G.; Kang, P.T.; Green-Church, K.B.; Chan, M.K.; Krzycki, J.A.
    Direct charging of tRNA(CUA) with pyrrolysine in vitro and in vivo (2004), Nature, 431, 333-335.
    View publication on PubMed

Cloned(Commentary)

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

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

Metals/Ions Comment Organism Structure
Mg2+
-
Methanosarcina barkeri

Natural Substrates/ Products (Substrates)

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

Organism UniProt Comment Textmining
Methanosarcina barkeri
-
gene pylS
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His6-tagged PylS from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Methanosarcina barkeri

Substrates and Products (Substrate)

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

Synonyms Comment Organism
class II aminoacyl-tRNA synthetase
-
Methanosarcina barkeri
PylS
-
Methanosarcina barkeri

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Methanosarcina barkeri

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.2
-
assay at Methanosarcina barkeri

Cofactor

Cofactor Comment Organism Structure
ATP
-
Methanosarcina barkeri