Cloned (Comment) | Organism |
---|---|
expression of the His-tagged CP1 domain mutant and isolated CP1 domain in Escherichia coli strain JM109(DE3) | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
additional information | construction of an enzyme mutant with a duplication of the peptide fragment from Met238 to Pro368 within the CP1 domain which shows an activity reduced by 59% compared to the wild-type enzyme, and catalyzes the mischarging of tRNALeu by methionine or isoleucine due to impaired ability to edit incorrect products | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0015 | - |
tRNALeu | aminoacylation reaction, wild-type enzyme, pH 7.8, 37°C | Escherichia coli | |
0.0024 | - |
tRNALeu | aminoacylation reaction, mutant enzyme, pH 7.8, 37°C | Escherichia coli | |
0.015 | - |
L-leucine | aminoacylation reaction, wild-type and mutant enzyme, pH 7.8, 37°C | Escherichia coli | |
0.052 | - |
L-leucine | ATP-diphosphate exchange reaction, wild-type enzyme, pH 7.8, 37°C | Escherichia coli | |
0.069 | - |
L-leucine | ATP-diphosphate exchange reaction, mutant enzyme, pH 7.8, 37°C | Escherichia coli | |
0.25 | - |
ATP | aminoacylation reaction, mutant enzyme, pH 7.8, 37°C | Escherichia coli | |
0.28 | - |
ATP | aminoacylation reaction, wild-type enzyme, pH 7.8, 37°C | Escherichia coli | |
2.8 | - |
L-isoleucine | ATP-diphosphate exchange reaction, mutant enzyme, pH 7.8, 37°C | Escherichia coli | |
3.5 | - |
L-isoleucine | ATP-diphosphate exchange reaction, wild-type enzyme, pH 7.8, 37°C | Escherichia coli | |
6.2 | - |
L-methionine | ATP-diphosphate exchange reaction, mutant enzyme, pH 7.8, 37°C | Escherichia coli | |
7.5 | - |
L-methionine | ATP-diphosphate exchange reaction, wild-type enzyme, pH 7.8, 37°C | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-leucine + tRNALeu | Escherichia coli | - |
AMP + diphosphate + L-leucyl-tRNALeu | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged mutant and His-tagged CP1 domain expressed in Escherichia coli strain Jm109(DE3), to electrophoretic homogeneity | Escherichia coli |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.79 | - |
recombinant CP1 domain mutant | Escherichia coli |
1.5 | - |
about, wild-type enzyme | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-isoleucine + tRNALeu | wild-type and CP1 domain mutant enzyme, the mischarged product can be edited by the wild-type enzyme, but not by a recombinant isolated CP1 domain | Escherichia coli | AMP + diphosphate + L-isoleucyl-tRNALeu | - |
? | |
ATP + L-leucine + tRNALeu | - |
Escherichia coli | AMP + diphosphate + L-leucyl-tRNALeu | - |
? | |
ATP + L-leucine + tRNALeu | two-step reaction, the connecting peptide CP1 domain is crucial for the editing function | Escherichia coli | AMP + diphosphate + L-leucyl-tRNALeu | - |
? | |
ATP + L-methionine + tRNALeu | wild-type and CP1 domain mutant enzyme, the mischarged product can be edited by the wild-type enzyme, but not by a recombinant isolated CP1 domain | Escherichia coli | AMP + diphosphate + L-methionyl-tRNALeu | - |
? | |
additional information | the enzyme also performs the ATP-diphosphate exchange reaction | Escherichia coli | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | determination of secondary structure elements in the wild-type and mutant enzyme | Escherichia coli |
Synonyms | Comment | Organism |
---|---|---|
Leucine translase | - |
Escherichia coli |
Leucine--tRNA ligase | - |
Escherichia coli |
Leucyl-transfer ribonucleate synthetase | - |
Escherichia coli |
Leucyl-transfer ribonucleic acid synthetase | - |
Escherichia coli |
Leucyl-transfer RNA synthetase | - |
Escherichia coli |
Leucyl-tRNA synthetase | - |
Escherichia coli |
LeuRS | - |
Escherichia coli |
Synthetase, leucyl-transfer ribonucleate | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.3 | - |
tRNALeu | mutant enzyme, pH 7.8, 37°C | Escherichia coli | |
1.5 | - |
L-leucine | mutant enzyme, pH 7.8, 37°C | Escherichia coli | |
1.6 | - |
ATP | mutant enzyme, pH 7.8, 37°C | Escherichia coli | |
2.9 | - |
tRNALeu | wild-type enzyme, pH 7.8, 37°C | Escherichia coli | |
3 | - |
L-leucine | wild-type enzyme, pH 7.8, 37°C | Escherichia coli | |
3.6 | - |
ATP | wild-type enzyme, pH 7.8, 37°C | Escherichia coli | |
6.9 | - |
L-isoleucine | ATP-diphosphate exchange reaction, mutant enzyme, pH 7.8, 37°C | Escherichia coli | |
7.6 | - |
L-methionine | ATP-diphosphate exchange reaction, mutant enzyme, pH 7.8, 37°C | Escherichia coli | |
18 | - |
L-isoleucine | ATP-diphosphate exchange reaction, wild-type enzyme, pH 7.8, 37°C | Escherichia coli | |
19 | - |
L-methionine | ATP-diphosphate exchange reaction, wild-type enzyme, pH 7.8, 37°C | Escherichia coli | |
101 | - |
L-leucine | ATP-diphosphate exchange reaction, mutant enzyme, pH 7.8, 37°C | Escherichia coli | |
171 | - |
L-leucine | ATP-diphosphate exchange reaction, wild-type enzyme, pH 7.8, 37°C | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.8 | - |
assay at | Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Escherichia coli |