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Literature summary extracted from

  • Sanford, B.; Cao, B.; Johnson, J.M.; Zimmerman, K.; Strom, A.M.; Mueller, R.M.; Bhattacharyya, S.; Musier-Forsyth, K.; Hati, S.
    Role of coupled dynamics in the catalytic activity of prokaryotic-like prolyl-tRNA synthetases (2012), Biochemistry, 51, 2146-2156.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.1.1.15 expressed in Escherichia coli SG13009 (pREP4) cells Escherichia coli

Protein Variants

EC Number Protein Variants Comment Organism
6.1.1.15 E218A the mutant activates proline but with a decreased kcat (3fold) and elevated KM value (15fold). Overall proline activation efficiency of this mutant is decreased 45fold compared to the wild type enzyme. The mutant also can charge L-proline onto tRNAPro, albeit with 3fold reduced efficiency Escherichia coli
6.1.1.15 G217A the kcat/KM of the mutant is reduced 7fold relative to the wild type enzyme. In contrast, alanine activation by the G217A mutant is not affected compared to the wild type enzyme. A 2fold decrease in alanine activation s observed for the mutant compared to the wild type enzyme. The mutant also can charge L-proline onto tRNAPro, albeit with 3fold reduced efficiency Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.1.1.15 0.228
-
 L-proline wild type enzyme, at pH 7.0 and 22°C Escherichia coli
6.1.1.15 0.427
-
 L-proline mutant enzyme G217A, at pH 7.0 and 22°C Escherichia coli
6.1.1.15 3.4
-
 L-proline mutant enzyme E218A, at pH 7.0 and 22°C Escherichia coli
6.1.1.15 454
-
 L-alanine mutant enzyme G217A, at pH 7.0 and 22°C Escherichia coli
6.1.1.15 685
-
 L-alanine wild type enzyme, at pH 7.0 and 22°C Escherichia coli
6.1.1.15 1360
-
 L-alanine mutant enzyme E218A, at pH 7.0 and 22°C Escherichia coli

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.1.1.15 ATP + L-proline + tRNAPro Escherichia coli
-
 AMP + diphosphate + L-prolyl-tRNAPro
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
6.1.1.15 Escherichia coli P16659
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.1.1.15 Talon cobalt affinity resin column chromatography Escherichia coli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.1.1.15 ATP + L-alanine + tRNAPro very low activity Escherichia coli AMP + diphosphate + L-alanyl-tRNAPro
-
 ?
6.1.1.15 ATP + L-proline + tRNAPro
-
 Escherichia coli AMP + diphosphate + L-prolyl-tRNAPro
-
 ?

Synonyms

EC Number Synonyms Comment Organism
6.1.1.15 Prolyl-tRNA synthetase
-
 Escherichia coli
6.1.1.15 ProRS
-
 Escherichia coli

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
6.1.1.15 2.18
-
 L-alanine mutant enzyme G217A, at pH 7.0 and 22°C Escherichia coli
6.1.1.15 3.26
-
 L-alanine mutant enzyme E218A, at pH 7.0 and 22°C Escherichia coli
6.1.1.15 3.37
-
 L-proline mutant enzyme G217A, at pH 7.0 and 22°C Escherichia coli
6.1.1.15 3.52
-
 L-alanine wild type enzyme, at pH 7.0 and 22°C Escherichia coli
6.1.1.15 4.4
-
 L-proline mutant enzyme E218A, at pH 7.0 and 22°C Escherichia coli
6.1.1.15 12.7
-
 L-proline wild type enzyme, at pH 7.0 and 22°C Escherichia coli

Cofactor

EC Number Cofactor Comment Organism Structure
6.1.1.15 ATP
-
 Escherichia coli

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
6.1.1.15 0.0024
-
 L-alanine mutant enzyme E218A, at pH 7.0 and 22°C Escherichia coli
6.1.1.15 0.0048
-
 L-alanine mutant enzyme G217A, at pH 7.0 and 22°C Escherichia coli
6.1.1.15 0.00513
-
 L-alanine wild type enzyme, at pH 7.0 and 22°C Escherichia coli
6.1.1.15 1.29
-
 L-proline mutant enzyme E218A, at pH 7.0 and 22°C Escherichia coli
6.1.1.15 7.89
-
 L-proline mutant enzyme G217A, at pH 7.0 and 22°C Escherichia coli
6.1.1.15 55.7
-
 L-proline wild type enzyme, at pH 7.0 and 22°C Escherichia coli