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Literature summary for 6.1.1.15 extracted from

  • Stathopoulos, C.; Jacquin-Becker, C.; Becker, H.D.; Li, T.; Ambrogelly, A.; Longman, R.; Soll, D.
    Methanococcus jannaschii prolyl-cysteinyl-tRNA synthetase possesses overlapping amino acid binding sites (2001), Biochemistry, 40, 46-52.
    View publication on PubMed
Search for more literature for 6.1.1.15

Cloned(Commentary)

Cloned (Comment) Organism
recombinant His-tagged wild-type and mutants from overexpressing Escherichia coli BL21 Methanocaldococcus jannaschii

Protein Variants

Protein Variants Comment Organism
E103A unaltered thermostability, no remaining prolylation activity, 5% remaining cysteinylation activity compared to the wild-type enzyme Methanocaldococcus jannaschii
P100A unaltered thermostability, loss of 90% cysteinylation activity, unaltered prolylation activity compared to the wild-type enzyme Methanocaldococcus jannaschii

Inhibitors

Inhibitors Comment Organism Structure
L-cysteine competitive inhibition of prolylation. A 40fold excess over L-proline concentration reduces the prolylation activity by 80%, no inhibition of mutant P100A Methanocaldococcus jannaschii
L-proline competitive inhibition of cysteinylation. A 40fold excess over L-cysteine concentration reduces the cysteinylation activity by over 80%, no inhibition of mutant E103A Methanocaldococcus jannaschii

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.022
-
 L-cysteine aminoacylation reaction, recombinant wild-type enzyme, pH 7.0, 70°C Methanocaldococcus jannaschii
0.027
-
 L-proline aminoacylation reaction, recombinant wild-type enzyme, pH 7.0, 70°C Methanocaldococcus jannaschii
0.049
-
 ATP aminoacylation reaction with L-proline, recombinant wild-type enzyme, pH 7.0, 70°C Methanocaldococcus jannaschii
0.06
-
 ATP aminoacylation reaction with L-cysteine, recombinant wild-type enzyme, pH 7.0, 70°C Methanocaldococcus jannaschii

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + L-proline + tRNAPro Methanocaldococcus jannaschii
-
 AMP + diphosphate + L-prolyl-tRNAPro
-
 ?
ATP + L-proline + tRNAPro Methanocaldococcus jannaschii DSM 2661
-
 AMP + diphosphate + L-prolyl-tRNAPro
-
 ?

Organism

Organism UniProt Comment Textmining
Methanocaldococcus jannaschii Q58635
-
-
Methanocaldococcus jannaschii DSM 2661 Q58635
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + L-cysteine + tRNA in later work it is shown that cysteine is attached to tRNA(Pro) and thus constitutes a misaminoacylation event and not a dual specificity Methanocaldococcus jannaschii AMP + diphosphate + L-cysteinyl-tRNA
-
 ?
ATP + L-cysteine + tRNA in later work it is shown that cysteine is attached to tRNA(Pro) and thus constitutes a misaminoacylation event and not a dual specificity Methanocaldococcus jannaschii DSM 2661 AMP + diphosphate + L-cysteinyl-tRNA
-
 ?
ATP + L-proline + tRNAPro
-
 Methanocaldococcus jannaschii AMP + diphosphate + L-prolyl-tRNAPro
-
 ?
ATP + L-proline + tRNAPro
-
 Methanocaldococcus jannaschii DSM 2661 AMP + diphosphate + L-prolyl-tRNAPro
-
 ?

Synonyms

Synonyms Comment Organism
ProCysRS misleading Methanocaldococcus jannaschii
proline/cysteine-tRNA ligase misleading Methanocaldococcus jannaschii
prolyl-cysteinyl-tRNA synthetase misleading Methanocaldococcus jannaschii

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
70
-
 assay at Methanocaldococcus jannaschii

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.02
-
 L-cysteine aminoacylation reaction, recombinant wild-type enzyme, pH 7.0, 70°C Methanocaldococcus jannaschii
0.09
-
 L-proline aminoacylation reaction, recombinant wild-type enzyme, pH 7.0, 70°C Methanocaldococcus jannaschii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
 assay at Methanocaldococcus jannaschii