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Literature summary for 6.1.1.26 extracted from

  • Herring, S.; Ambrogelly, A.; Gundllapalli, S.; O'Donoghue, P.; Polycarpo, C.R.; Soll, D.
    The amino-terminal domain of pyrrolysyl-tRNA synthetase is dispensable in vitro but required for in vivo activity (2007), FEBS Lett., 581, 3197-3203.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 6.1.1.26

Cloned(Commentary)

Cloned (Comment) Organism
gene pylS, DNA and amino acid sequence determination and analysis, expression in Escherichia coli BL21(DE3) as His-tagged enzyme Methanosarcina thermophila
gene pylS, expression in Escherichia coli BL21(DE3) as His-tagged enzyme Desulfitobacterium hafniense
gene pylS, expression of His-tagged wild-type and mutant enzymes in Escherichia coli BL21(DE3) Methanosarcina barkeri

Protein Variants

Protein Variants Comment Organism
D2A site-directed mutagenesis, the mutant shows 95% reduced activity compared to the wild-type enzyme Methanosarcina barkeri
D2A/K4A site-directed mutagenesis, the mutant shows almost completely reduced activity compared to the wild-type enzyme Methanosarcina barkeri
D7A site-directed mutagenesis, the mutant shows only slightly reduced activity compared to the wild-type enzyme Methanosarcina barkeri
G21L site-directed mutagenesis, the mutant shows only slightly reduced activity compared to the wild-type enzyme Methanosarcina barkeri
H24A site-directed mutagenesis, the mutant shows 95% reduced activity compared to the wild-type enzyme Methanosarcina barkeri
I26G site-directed mutagenesis, the mutant shows 80% reduced activity compared to the wild-type enzyme Methanosarcina barkeri
K3A site-directed mutagenesis, the mutant shows 80% reduced activity compared to the wild-type enzyme Methanosarcina barkeri
K4A site-directed mutagenesis, the mutant shows 95% reduced activity compared to the wild-type enzyme Methanosarcina barkeri
additional information construction of several truncation mutants, which show 90-99% reduced activity compared to the wild-type enzyme, overview Methanosarcina barkeri
R19A site-directed mutagenesis, the mutant shows only slightly reduced activity compared to the wild-type enzyme Methanosarcina barkeri
S11A site-directed mutagenesis, the mutant shows 95% reduced activity compared to the wild-type enzyme Methanosarcina barkeri
S11A/T13A site-directed mutagenesis, the mutant shows almost completely reduced activity compared to the wild-type enzyme Methanosarcina barkeri
S18A site-directed mutagenesis, the mutant shows only slightly reduced activity compared to the wild-type enzyme Methanosarcina barkeri
T13A site-directed mutagenesis, the mutant shows 95% reduced activity compared to the wild-type enzyme Methanosarcina barkeri
W16A site-directed mutagenesis, the mutant shows 35% reduced activity compared to the wild-type enzyme Methanosarcina barkeri

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00015 0.00098 additional information equilibrium binding analysis, dissociation constants of the enzyme with tRNAPyl from different species, overview Methanosarcina barkeri
0.00016 0.001 additional information equilibrium binding analysis, dissociation constants of the enzyme with tRNAPyl from different species, overview Methanosarcina thermophila
0.0053 0.0069 additional information equilibrium binding analysis, dissociation constants of the enzyme with tRNAPyl from different species, overview Desulfitobacterium hafniense

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+
-
 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
-
 ?
ATP + L-pyrrolysine + tRNAPyl Methanosarcina thermophila
-
 AMP + diphosphate + L-pyrrolysyl-tRNAPyl
-
 ?
additional information Methanosarcina thermophila the amino-terminal extension present in archaeal PylRSs is dispensable for in vitro activity, but required for PylRS function in vivo ?
-
 ?

Organism

Organism UniProt Comment Textmining
Desulfitobacterium hafniense
-
 a strictly anaerobic bacterium
-
Methanosarcina barkeri
-
 strains Fusaro and MS, gene pylS
-
Methanosarcina thermophila Q1L6A3 gene pylS
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme from Escherichia coli BL21(DE3) Desulfitobacterium hafniense
recombinant His-tagged enzyme from Escherichia coli BL21(DE3) Methanosarcina thermophila
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli BL21(DE3) Methanosarcina barkeri

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
-
 Methanosarcina thermophila AMP + diphosphate + L-pyrrolysyl-tRNAPyl
-
 ?
ATP + L-pyrrolysine + tRNAPyl the archaeal enzyme does not distinguish between archaeal and bacterial tRNAPyl species, substrate specificity, overview Methanosarcina thermophila AMP + diphosphate + L-pyrrolysyl-tRNAPyl
-
 ?
ATP + L-pyrrolysine + tRNAPyl the archaeal enzyme does not distinguish between archaeal and bacterial tRNAPyl species, substrate specificity, overview, residues from the PylRS amino-terminal domain affect activity in vivo Methanosarcina barkeri AMP + diphosphate + L-pyrrolysyl-tRNAPyl
-
 ?
ATP + L-pyrrolysine + tRNAPyl the bacterial PylRS displays a clear preference for the homologous cognate tRNA, substrate specificity, overview Desulfitobacterium hafniense AMP + diphosphate + L-pyrrolysyl-tRNAPyl
-
 ?
additional information the amino-terminal extension present in archaeal PylRSs is dispensable for in vitro activity, but required for PylRS function in vivo Methanosarcina thermophila ?
-
 ?

Subunits

Subunits Comment Organism
More the PylRS sequence can be subdivided into three regions: the highly conserved class II aaRS catalytic core domain at the carboxy-terminal, the unique amino-terminal domain, and a highly variable region linking these two domains Desulfitobacterium hafniense
More the PylRS sequence can be subdivided into three regions: the highly conserved class II aaRS catalytic core domain at the carboxy-terminal, the unique amino-terminal domain, and a highly variable region linking these two domains, residues from the PylRS amino-terminal domain affect activity in vivo Methanosarcina barkeri
More the PylRS sequence can be subdivided into three regions: the highly conserved class II aaRS catalytic core domain at the carboxy-terminal, the unique amino-terminal domain, and a highly variable region linking these two domains, the amino-terminal extension present in archaeal PylRSs is dispensable for in vitro activity, but required for PylRS function in vivo Methanosarcina thermophila

Synonyms

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

Temperature Optimum [°C]

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

pH Optimum

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

Cofactor

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