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Literature summary extracted from

  • Chen, X.; Ma, J.J.; Tan, M.; Yao, P.; Hu, Q.H.; Eriani, G.; Wang, E.D.
    Modular pathways for editing non-cognate amino acids by human cytoplasmic leucyl-tRNA synthetase (2011), Nucleic Acids Res., 39, 235-247.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.1.1.4 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

EC Number Protein Variants Comment Organism
6.1.1.4 D399A site-directed mutagenesis, tRNA selectivity compared to the wild-type enzyme Homo sapiens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.1.1.4 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
6.1.1.4 0.0456
-
 L-leucine pH 7.6, 37°C, wild-type enzyme Homo sapiens
6.1.1.4 0.05
-
 L-leucine pH 7.6, 37°C, mutant D399A Homo sapiens

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
6.1.1.4 cytoplasm
-
 Homo sapiens 5737
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
6.1.1.4 Mg2+ required Homo sapiens

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
6.1.1.4 135000
-
 gel filtration Homo sapiens

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.1.1.4 ATP + L-leucine + tRNALeu Homo sapiens
-
 AMP + diphosphate + L-leucyl-tRNALeu
-
 ?
6.1.1.4 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

EC Number Organism UniProt Comment Textmining
6.1.1.4 Homo sapiens
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.1.1.4 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 and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.1.1.4 ATP + L-leucine + tRNALeu
-
 Homo sapiens AMP + diphosphate + L-leucyl-tRNALeu
-
 ?
6.1.1.4 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

EC Number Synonyms Comment Organism
6.1.1.4 Leucyl-tRNA synthetase
-
 Homo sapiens
6.1.1.4 LeuRS
-
 Homo sapiens
6.1.1.4 More the enzyme belongs to the class Ia aminoacyl-tRNA transferases Homo sapiens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
6.1.1.4 37
-
 assay at Homo sapiens

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
6.1.1.4 25.8
-
 L-leucine pH 7.6, 37°C, wild-type enzyme Homo sapiens
6.1.1.4 26.2
-
 L-leucine pH 7.6, 37°C, mutant D399A Homo sapiens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
6.1.1.4 7.6
-
 assay at Homo sapiens

Cofactor

EC Number Cofactor Comment Organism Structure
6.1.1.4 ATP
-
 Homo sapiens

General Information

EC Number General Information Comment Organism
6.1.1.4 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 [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
6.1.1.4 524
-
 L-leucine pH 7.6, 37°C, mutant D399A Homo sapiens
6.1.1.4 565
-
 L-leucine pH 7.6, 37°C, wild-type enzyme Homo sapiens