Cloned (Comment) | Organism |
---|---|
gene thrS, sequence comparisons, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
E458D | site-directed mutagenesis, the sensitivity of the mutant enzyme to borrelidin is reduced markedly compared to wild-type, mutant shows decreased apparent rate constants | Escherichia coli |
G459D | site-directed mutagenesis, the sensitivity of the mutant enzyme to borrelidin is reduced markedly compared to wild-type, mutant shows decreased apparent rate constants | Escherichia coli |
P424K | site-directed mutagenesis, the sensitivity of the mutant enzyme to borrelidin is reduced markedly compared to wild-type, the mutant shows decreased apparent rate constants | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
borrelidin | is an 18-membered macrolide polyketide produced by several actinomycete bacteria of the Streptomyces spp.. Identification of borrelidin binding site on threonyl-tRNA synthetase, molecular docking, overview. Borrelidin binds the pocket outside but adjacent to the active site of ThrRS, consisting of residues Y313, R363, R375, P424, E458, G459, and K465. Borrelidin may induce the cleft closure, which blocks the release of Thr-AMP and phosphate, to inhibit activity of ThrRS rather than inhibit the binding of ATP and threonine | Escherichia coli | |
additional information | development of novel borrelidin derivatives and rational design of structure-based ThrRS inhibitors, overview | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | enzyme kinetics and stopped-flow fluorescence analysis, Michaelis-Menten steady-state kinetics of recombinant wild-type and mutant enzymes | Escherichia coli | |
0.101 | - |
L-threonine | pH 7.4, 30°C, recombinant mutant G459D | Escherichia coli | |
0.104 | - |
L-threonine | pH 7.4, 30°C, recombinant mutant E458D | Escherichia coli | |
0.105 | - |
L-threonine | pH 7.4, 30°C, recombinant wild-type enzyme | Escherichia coli | |
0.108 | - |
L-threonine | pH 7.4, 30°C, recombinant mutant P424K | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-threonine + tRNAThr | Escherichia coli | - |
AMP + diphosphate + L-threonyl-tRNAThr | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P0A8M3 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-threonine + tRNAThr | - |
Escherichia coli | AMP + diphosphate + L-threonyl-tRNAThr | - |
? |
Synonyms | Comment | Organism |
---|---|---|
Threonyl-tRNA synthetase | - |
Escherichia coli |
ThrRS | - |
Escherichia coli |
ThrS | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2 | 8 | L-threonine | pH 7.4, 30°C, recombinant mutant G459D | Escherichia coli | |
30 | - |
L-threonine | pH 7.4, 30°C, recombinant mutant P424K | Escherichia coli | |
34 | - |
L-threonine | pH 7.4, 30°C, recombinant mutant E458D | Escherichia coli | |
35 | - |
L-threonine | pH 7.4, 30°C, recombinant wild-type enzyme | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | - |
assay at | Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | binding structure | Escherichia coli |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0000034 | - |
borrelidin | pH 7.4, 30°C, recombinant wild-type enzyme | Escherichia coli | |
0.0000191 | - |
borrelidin | pH 7.4, 30°C, recombinant mutant E458D | Escherichia coli | |
0.0000651 | - |
borrelidin | pH 7.4, 30°C, recombinant mutant P424K | Escherichia coli | |
0.0001114 | - |
borrelidin | pH 7.4, 30°C, recombinant mutant G459D | Escherichia coli |