Cloned (Comment) | Organism |
---|---|
expression of His-tagged cytoplasmic LeuRS in Escherichia coli strain RosettaTM 2 (DE3), expression in Spodotera frugiperda cells via baculovirus transfection system | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
D399A | site-directed mutagenesis, tRNA selectivity compared to the wild-type enzyme | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetic constants of wild-type and D399A mutant of LeuRS in amino acid activation reaction with different amino acids, overview. Cytoplasmic LeuRS overexpressed in Escherichia coli exhibits the same kcat values as the one overexpressed in insect cells using in vitro transcribed tRNA | Homo sapiens | |
0.0456 | - |
L-leucine | pH 7.6, 37°C, wild-type enzyme | Homo sapiens | |
0.05 | - |
L-leucine | pH 7.6, 37°C, mutant D399A | Homo sapiens |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
cytoplasm | - |
Homo sapiens | 5737 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Homo sapiens |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
135000 | - |
gel filtration | Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-leucine + tRNALeu | Homo sapiens | - |
AMP + diphosphate + L-leucyl-tRNALeu | - |
? | |
additional information | Homo sapiens | LeuRS misactivates several non-cognate amino acids, e.g. Ile and Met as well as the non-standard amino acids norvaline and alpha-amino butyrate. It uses mainly pre-transfer editing to edit alpha-amino butyrate and a tRNA-dependent mechanism to edit norvaline, although both amino acids can be charged to tRNALeu, overview. Separation of the norvaline-editing pathways | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged LeuRS from Escherichia coli strain RosettaTM 2 (DE3) by nickel affinity and anion exchange chromatography, and ethanol precipitation, to 90% purity | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-leucine + tRNALeu | - |
Homo sapiens | AMP + diphosphate + L-leucyl-tRNALeu | - |
? | |
additional information | LeuRS misactivates several non-cognate amino acids, e.g. Ile and Met as well as the non-standard amino acids norvaline and alpha-amino butyrate. It uses mainly pre-transfer editing to edit alpha-amino butyrate and a tRNA-dependent mechanism to edit norvaline, although both amino acids can be charged to tRNALeu, overview. Separation of the norvaline-editing pathways | Homo sapiens | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
Leucyl-tRNA synthetase | - |
Homo sapiens |
LeuRS | - |
Homo sapiens |
More | the enzyme belongs to the class Ia aminoacyl-tRNA transferases | Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Homo sapiens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
25.8 | - |
L-leucine | pH 7.6, 37°C, wild-type enzyme | Homo sapiens | |
26.2 | - |
L-leucine | pH 7.6, 37°C, mutant D399A | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.6 | - |
assay at | Homo sapiens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Homo sapiens |
General Information | Comment | Organism |
---|---|---|
physiological function | aminoacyl-tRNA synthetases have evolved editing mechanisms to hydrolyze misactivated amino acids (pre-transfer editing) or misacylated tRNAs (post-transfer editing). Class Ia leucyl-tRNA synthetase may misactivate various natural and non-protein amino acids and then mischarge tRNALeu. The fidelity of prokaryotic LeuRS depends on multiple editing pathways to clear the incorrect intermediates and products in every step of aminoacylation reaction. Post-transfer editing as a final checkpoint of the reaction is very important to prevent mis-incorporation in vitro | Homo sapiens |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
524 | - |
L-leucine | pH 7.6, 37°C, mutant D399A | Homo sapiens | |
565 | - |
L-leucine | pH 7.6, 37°C, wild-type enzyme | Homo sapiens |