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Literature summary for 6.1.1.4 extracted from

  • Tan, M.; Zhu, B.; Liu, R.J.; Chen, X.; Zhou, X.L.; Wang, E.D.
    Interdomain communication modulates the tRNA-dependent pre-transfer editing of leucyl-tRNA synthetase (2013), Biochem. J., 449, 123-131.
    View publication on PubMed
Search for more literature for 6.1.1.4

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Escherichia coli

Protein Variants

Protein Variants Comment Organism
E184A the mutant performs the activities of amino acid activation, aminoacylation and deacylation of mischarged tRNAs as well as the native enzyme Escherichia coli
E184R the mutant performs the activities of amino acid activation, aminoacylation and deacylation of mischarged tRNAs as well as the native enzyme. The substitution specifically inhibits tRNA-dependent pre-transfer editing Escherichia coli
E184R/T252R the mutant performs the activities of amino acid activation, aminoacylation and deacylation of mischarged tRNAs as well as the native enzyme Escherichia coli
R185E the mutation significantly enhances tRNA-dependent pre-transfer editing activity Escherichia coli
R286E the mutation significantly enhances tRNA-dependent pre-transfer editing activity Escherichia coli
T252R the mutant performs the activities of amino acid activation, aminoacylation and deacylation of mischarged tRNAs as well as the native enzyme Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.002
-
 Ile-tRNALeu mutant enzyme R185E, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C Escherichia coli
0.0021
-
 Ile-tRNALeu wild type enzyme, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C Escherichia coli
0.0024
-
 Ile-tRNALeu mutant enzyme R286E, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C Escherichia coli
0.0025
-
 Ile-tRNALeu mutant enzyme E184R, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C Escherichia coli
0.039
-
 L-leucine wild type enzyme, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C 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
-
 ?
additional information Escherichia coli the enzyme has evolved both tRNA-dependent pre- and post-transfer editing capabilities to ensure catalytic specificity ?
-
 ?

Organism

Organism UniProt Comment Textmining
Escherichia coli P07813
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA column chromatography Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
AMP + diphosphate + Ile-tRNALeu
-
 Escherichia coli ATP + L-isoleucine + tRNALeu
-
 ?
ATP + L-leucine + tRNALeu
-
 Escherichia coli AMP + diphosphate + L-leucyl-tRNALeu
-
 ?
additional information the enzyme has evolved both tRNA-dependent pre- and post-transfer editing capabilities to ensure catalytic specificity Escherichia coli ?
-
 ?

Synonyms

Synonyms Comment Organism
Leucyl-tRNA synthetase
-
 Escherichia coli
LeuRS
-
 Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
7
-
 Ile-tRNALeu wild type enzyme, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C Escherichia coli
7.4
-
 Ile-tRNALeu mutant enzyme R286E, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C Escherichia coli
7.6
-
 Ile-tRNALeu mutant enzyme R185E, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C Escherichia coli
8.8
-
 Ile-tRNALeu mutant enzyme E184R, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C Escherichia coli
73.7
-
 L-leucine wild type enzyme, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C Escherichia coli

Cofactor

Cofactor Comment Organism Structure
ATP
-
 Escherichia coli

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1890
-
 L-leucine wild type enzyme, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C Escherichia coli
3100
-
 Ile-tRNALeu mutant enzyme R286E, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C Escherichia coli
3300
-
 Ile-tRNALeu wild type enzyme, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C Escherichia coli
3500
-
 Ile-tRNALeu mutant enzyme E184R, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C Escherichia coli
3800
-
 Ile-tRNALeu mutant enzyme R185E, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C Escherichia coli