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Literature summary for 6.1.1.15 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
Search for more literature for 6.1.1.15

Cloned(Commentary)

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

Protein Variants

Protein Variants Comment Organism
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
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]

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

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Escherichia coli P16659
-
-

Purification (Commentary)

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

Substrates and Products (Substrate)

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

Synonyms

Synonyms Comment Organism
Prolyl-tRNA synthetase
-
 Escherichia coli
ProRS
-
 Escherichia coli

Turnover Number [1/s]

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

Cofactor

Cofactor Comment Organism Structure
ATP
-
 Escherichia coli

kcat/KM [mM/s]

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