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Literature summary for 6.1.1.26 extracted from

  • Jiang, R.; Krzycki, J.A.
    PylSn and the homologous N-terminal domain of pyrrolysyl-tRNA synthetase bind the tRNA that is essential for the genetic encoding of pyrrolysine (2012), J. Biol. Chem., 287, 32738-32746.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 6.1.1.26

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Methanosarcina barkeri
expressed in Escherichia coli BL21(DE3) cells Desulfitobacterium hafniense

Protein Variants

Protein Variants Comment Organism
additional information truncation of the N-terminal region of PylS (DELTA92PylS) eliminates detectable tRNAPyl binding, but not catalytic activity Methanosarcina barkeri

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.044
-
 L-pyrrolysine wild type enzyme, at 37°C in 50 mM KCl, 10 mM MgCl2, 5 mM ATP, and 5 mM dithiothreitol in 10 mM HEPES buffer, pH 7.2 Methanosarcina barkeri
0.044
-
 L-pyrrolysine wild type enzyme, at 37°C in 50 mM KCl, 10 mM MgCl2, 5 mM ATP, and 5 mM dithiothreitol in 10 mM HEPES buffer, pH 7.2 Desulfitobacterium hafniense

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + L-pyrrolysine + tRNAPyl Methanosarcina barkeri
-
 AMP + diphosphate + L-pyrrolysyl-tRNAPyl
-
 ?
ATP + L-pyrrolysine + tRNAPyl Desulfitobacterium hafniense
-
 AMP + diphosphate + L-pyrrolysyl-tRNAPyl
-
 ?

Organism

Organism UniProt Comment Textmining
Desulfitobacterium hafniense
-
-
-
Methanosarcina barkeri
-
-
-
Methanosarcina barkeri Q6WRH6
-
-

Purification (Commentary)

Purification (Comment) Organism
nickel-activated HiTrap column chromatography Methanosarcina barkeri
nickel-activated HiTrap column chromatography Desulfitobacterium hafniense

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + L-pyrrolysine + tRNAPyl
-
 Methanosarcina barkeri AMP + diphosphate + L-pyrrolysyl-tRNAPyl
-
 ?
ATP + L-pyrrolysine + tRNAPyl
-
 Desulfitobacterium hafniense AMP + diphosphate + L-pyrrolysyl-tRNAPyl
-
 ?
ATP + L-pyrrolysine + tRNAPyl the N-terminal region of PylS influences complex formation with tRNAPyl Methanosarcina barkeri AMP + diphosphate + L-pyrrolysyl-tRNAPyl
-
 ?

Synonyms

Synonyms Comment Organism
PylRS
-
 Methanosarcina barkeri
PylRS
-
 Desulfitobacterium hafniense
PylS
-
 Methanosarcina barkeri
PylS
-
 Desulfitobacterium hafniense
PylSn
-
 Methanosarcina barkeri
pyrrolysyl-tRNA synthetase
-
 Methanosarcina barkeri
pyrrolysyl-tRNA synthetase
-
 Desulfitobacterium hafniense

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.1
-
 L-pyrrolysine wild type enzyme, at 37°C in 50 mM KCl, 10 mM MgCl2, 5 mM ATP, and 5 mM dithiothreitol in 10 mM HEPES buffer, pH 7.2 Methanosarcina barkeri
0.325
-
 L-pyrrolysine wild type enzyme, at 37°C in 50 mM KCl, 10 mM MgCl2, 5 mM ATP, and 5 mM dithiothreitol in 10 mM HEPES buffer, pH 7.2 Methanosarcina barkeri
0.325
-
 L-pyrrolysine wild type enzyme, at 37°C in 50 mM KCl, 10 mM MgCl2, 5 mM ATP, and 5 mM dithiothreitol in 10 mM HEPES buffer, pH 7.2 Desulfitobacterium hafniense

Cofactor

Cofactor Comment Organism Structure
ATP
-
 Methanosarcina barkeri
ATP
-
 Desulfitobacterium hafniense