Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli SG13009 (pREP4) cells | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
D198A | the overall aminoacylation activity of the mutant is reduced 5.5fold | Escherichia coli |
E218A/N305A | inactive | Escherichia coli |
E234A | the overall aminoacylation activity of the mutant is reduced 2fold | Escherichia coli |
F415A | the aminoacylation activity of the mutant is nearly abolished with rates 70fold slower than the wild type | Escherichia coli |
G412A | the overall aminoacylation activity of the mutant is reduced 7fold | Escherichia coli |
H302A | the overall aminoacylation activity of the mutant is reduced 2fold | Escherichia coli |
H302A/G412A | the overall aminoacylation activity of the mutant is reduced 5.5fold | Escherichia coli |
N305A | the aminoacylation activity of the mutant is nearly abolished with rates 70fold slower than the wild type | Escherichia coli |
N305A/G412A | inactive | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.18 | - |
L-proline | wild type enzyme, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C | Escherichia coli | |
0.22 | - |
L-proline | mutant enzyme H302A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C | Escherichia coli | |
0.3 | - |
L-proline | mutant enzyme G412A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C | Escherichia coli | |
0.33 | - |
L-proline | mutant enzyme D198A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C | Escherichia coli | |
0.45 | - |
L-proline | mutant enzyme N305A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C | Escherichia coli | |
0.62 | - |
L-proline | mutant enzyme H302A/G412A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C | Escherichia coli | |
0.76 | - |
L-proline | mutant enzyme F415A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C | Escherichia coli | |
1.03 | - |
L-proline | mutant enzyme E234A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C | Escherichia coli | |
3.4 | - |
L-proline | mutant enzyme E218A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-proline + tRNAPro | Escherichia coli | - |
AMP + diphosphate + L-prolyl-tRNAPro | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Purification (Comment) | Organism |
---|---|
Talon cobalt affinity resin column chromatography, and gel filtration | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-proline + tRNAPro | - |
Escherichia coli | AMP + diphosphate + L-prolyl-tRNAPro | - |
r |
Synonyms | Comment | Organism |
---|---|---|
Prolyl-tRNA synthetase | - |
Escherichia coli |
ProRS | - |
Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.131 | - |
L-proline | mutant enzyme F415A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C | Escherichia coli | |
0.61 | - |
L-proline | mutant enzyme N305A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C | Escherichia coli | |
4.4 | - |
L-proline | mutant enzyme E218A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C | Escherichia coli | |
6.7 | - |
L-proline | mutant enzyme E234A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C | Escherichia coli | |
6.98 | - |
L-proline | mutant enzyme D198A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C | Escherichia coli | |
7.3 | - |
L-proline | mutant enzyme H302A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C | Escherichia coli | |
10.7 | - |
L-proline | mutant enzyme H302A/G412A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C | Escherichia coli | |
12.6 | - |
L-proline | wild type enzyme, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C | Escherichia coli | |
12.8 | - |
L-proline | mutant enzyme G412A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C | Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Escherichia coli |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.17 | - |
L-proline | mutant enzyme F415A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C | Escherichia coli | |
1.3 | - |
L-proline | mutant enzyme E218A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C | Escherichia coli | |
1.4 | - |
L-proline | mutant enzyme N305A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C | Escherichia coli | |
6.5 | - |
L-proline | mutant enzyme E234A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C | Escherichia coli | |
17 | - |
L-proline | mutant enzyme H302A/G412A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C | Escherichia coli | |
21 | - |
L-proline | mutant enzyme D198A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C | Escherichia coli | |
33 | - |
L-proline | mutant enzyme H302A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C | Escherichia coli | |
43 | - |
L-proline | mutant enzyme G412A, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C | Escherichia coli | |
71 | - |
L-proline | wild type enzyme, in 50 mM HEPES (pH 7.5), 20 mM KCl, 25 mM MgCl2, at 37°C | Escherichia coli |