Any feedback?
Literature summary for 6.1.1.4 extracted from
- Effect of alanine-293 replacement on the activity, ATP binding, and editing of Escherichia coli leucyl-tRNA synthetase (2001), Biochemistry, 40, 1144-1149.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expression of wild-type and mutants in Escherichia coli strain TG1 | Escherichia coli |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| A293D | 81% decreased activity, highly decreased editing function, very strong binding of ATP, high decrease in Km for the substrates, more sensitive too inhibition by ATP, increased heat lability compared to the wild-type | Escherichia coli |
| A293F | 54% decreased activity compared to the wild-type, more sensitive too inhibition by ATP | Escherichia coli |
| A293G | 50% decreased activity, decreased editing function, stronger binding of ATP, decrease in Km for the substrates, more sensitive too inhibition by ATP | Escherichia coli |
| A293I | 51% decreased activity, decreased editing function, stronger binding of ATP, decrease in Km for the substrates, more sensitive too inhibition by ATP | Escherichia coli |
| A293R | 30% decreased activity, decreased editing function, stronger binding of ATP, decrease in Km for the substrates | Escherichia coli |
| A293Y | 50% decreased activity, decreased editing function, stronger binding of ATP, decrease in Km for the substrates, more sensitive too inhibition by ATP | Escherichia coli |
| M328K | 7% increased activity compared to the wild-type | Escherichia coli |
| additional information | construction of several CP1 domain mutants by introduction of restriction endonuclease sites into gene leuS | Escherichia coli |
| T252E | activity is similar to the wild-type | Escherichia coli |
| T252E/M328K | activity is similar to the wild-type | Escherichia coli |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| ATP | at high concentration, the A293 mutants are ore sensitive | Escherichia coli |  |
Natural Substrates/ Products (Substrates)
| 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
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Escherichia coli | - |
class I enzyme | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant wild-type and mutant enzymes | Escherichia coli |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| ATP + L-leucine + tRNALeu = AMP + diphosphate + L-leucyl-tRNALeu | A293 is important for the stability of the enzyme conformation and the editing function and probably is involved in the ATP binding | Escherichia coli |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 0.024 | - |
mutant A293D, in crude enzyme extract of Escherichia coli strain TG1 | Escherichia coli |
| 0.13 | - |
mutant A293F, in crude enzyme extract of Escherichia coli strain TG1 | Escherichia coli |
| 0.14 | - |
mutants A293I, A293G, and A293Y, in crude enzyme extract of Escherichia coli strain TG1 | Escherichia coli |
| 0.2 | - |
mutant A293R, in crude enzyme extract of Escherichia coli strain TG1 | Escherichia coli |
| 0.27 | - |
mutant T252E, in crude enzyme extract of Escherichia coli strain TG1 | Escherichia coli |
| 0.28 | - |
wild-type enzyme, in crude enzyme extract of Escherichia coli strain TG1 | Escherichia coli |
| 0.29 | - |
mutant T252E/M328K, in crude enzyme extract of Escherichia coli strain TG1 | Escherichia coli |
| 0.3 | - |
mutant M328K, in crude enzyme extract of Escherichia coli strain TG1 | Escherichia coli |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ATP + L-leucine + tRNALeu | - |
Escherichia coli | AMP + diphosphate + L-leucyl-tRNALeu | - |
? | |
| ATP + L-leucine + tRNALeu | the connecting peptide CP1 domain is crucial for the editing function | Escherichia coli | AMP + diphosphate + L-leucyl-tRNALeu | - |
? | |
Synonyms
| 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 [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | - |
assay at | Escherichia coli |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 49.5 | - |
50% inactivation, heating of 0.5°C per minute starting at 30°C, mutant A293D | Escherichia coli |
| 54 | - |
50% inactivation, heating of 0.5°C per minute starting at 30°C, wild-type enzyme and mutants A293F, A293G, A293R | Escherichia coli |
| 55 | - |
50% inactivation, heating of 0.5°C per minute starting at 30°C, mutants A293I and A293Y | Escherichia coli |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | kinetics, mutant enzymes | Escherichia coli | |
| 5 | - |
ATP | wild-type enzyme, pH 7.8, 37°C | Escherichia coli | |
| 5.1 | - |
tRNALeu | wild-type enzyme, pH 7.8, 37°C | Escherichia coli | |
| 5.2 | - |
L-leucine | wild-type enzyme, pH 7.8, 37°C | Escherichia coli | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.8 | - |
assay at | Escherichia coli |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| ATP | optimal concentration is 4 mM for the wild-type and mutant A293R, for the mutants A293Y, A293G, and A293I it is 2 mM, for the mutants A293D, and As93F it is 1 mM | Escherichia coli | |
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2023.1 (January 2023)
Legal
Curated at
Member of
