BRENDA - Enzyme Database

Interdomain communication modulates the tRNA-dependent pre-transfer editing of leucyl-tRNA synthetase

Tan, M.; Zhu, B.; Liu, R.J.; Chen, X.; Zhou, X.L.; Wang, E.D.; Biochem. J. 449, 123-131 (2013) View publication on PubMed

Data extracted from this reference:

Cloned(Commentary)
EC Number
Cloned (Commentary)
Organism
6.1.1.4
expressed in Escherichia coli BL21(DE3) cells
Escherichia coli
Engineering
EC Number
Protein Variants
Commentary
Organism
6.1.1.4
E184A
the mutant performs the activities of amino acid activation, aminoacylation and deacylation of mischarged tRNAs as well as the native enzyme
Escherichia coli
6.1.1.4
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
6.1.1.4
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
6.1.1.4
R185E
the mutation significantly enhances tRNA-dependent pre-transfer editing activity
Escherichia coli
6.1.1.4
R286E
the mutation significantly enhances tRNA-dependent pre-transfer editing activity
Escherichia coli
6.1.1.4
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]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
6.1.1.4
0.002
-
Ile-tRNALeu
mutant enzyme R185E, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
Escherichia coli
6.1.1.4
0.0021
-
Ile-tRNALeu
wild type enzyme, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
Escherichia coli
6.1.1.4
0.0024
-
Ile-tRNALeu
mutant enzyme R286E, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
Escherichia coli
6.1.1.4
0.0025
-
Ile-tRNALeu
mutant enzyme E184R, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
Escherichia coli
6.1.1.4
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)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
6.1.1.4
ATP + L-leucine + tRNALeu
Escherichia coli
-
AMP + diphosphate + L-leucyl-tRNALeu
-
-
?
6.1.1.4
additional information
Escherichia coli
the enzyme has evolved both tRNA-dependent pre- and post-transfer editing capabilities to ensure catalytic specificity
?
-
-
?
Organism
EC Number
Organism
UniProt
Commentary
Textmining
6.1.1.4
Escherichia coli
P07813
-
-
Purification (Commentary)
EC Number
Purification (Commentary)
Organism
6.1.1.4
Ni-NTA column chromatography
Escherichia coli
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
6.1.1.4
AMP + diphosphate + Ile-tRNALeu
-
726909
Escherichia coli
ATP + L-isoleucine + tRNALeu
-
-
-
?
6.1.1.4
ATP + L-leucine + tRNALeu
-
726909
Escherichia coli
AMP + diphosphate + L-leucyl-tRNALeu
-
-
-
?
6.1.1.4
additional information
the enzyme has evolved both tRNA-dependent pre- and post-transfer editing capabilities to ensure catalytic specificity
726909
Escherichia coli
?
-
-
-
?
Synonyms
EC Number
Synonyms
Commentary
Organism
6.1.1.4
Leucyl-tRNA synthetase
-
Escherichia coli
6.1.1.4
LeuRS
-
Escherichia coli
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
6.1.1.4
7
-
Ile-tRNALeu
wild type enzyme, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
Escherichia coli
6.1.1.4
7.4
-
Ile-tRNALeu
mutant enzyme R286E, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
Escherichia coli
6.1.1.4
7.6
-
Ile-tRNALeu
mutant enzyme R185E, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
Escherichia coli
6.1.1.4
8.8
-
Ile-tRNALeu
mutant enzyme E184R, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
Escherichia coli
6.1.1.4
73.7
-
L-leucine
wild type enzyme, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
Escherichia coli
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
6.1.1.4
ATP
-
Escherichia coli
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
6.1.1.4
expressed in Escherichia coli BL21(DE3) cells
Escherichia coli
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
6.1.1.4
ATP
-
Escherichia coli
Engineering (protein specific)
EC Number
Protein Variants
Commentary
Organism
6.1.1.4
E184A
the mutant performs the activities of amino acid activation, aminoacylation and deacylation of mischarged tRNAs as well as the native enzyme
Escherichia coli
6.1.1.4
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
6.1.1.4
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
6.1.1.4
R185E
the mutation significantly enhances tRNA-dependent pre-transfer editing activity
Escherichia coli
6.1.1.4
R286E
the mutation significantly enhances tRNA-dependent pre-transfer editing activity
Escherichia coli
6.1.1.4
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] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
6.1.1.4
0.002
-
Ile-tRNALeu
mutant enzyme R185E, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
Escherichia coli
6.1.1.4
0.0021
-
Ile-tRNALeu
wild type enzyme, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
Escherichia coli
6.1.1.4
0.0024
-
Ile-tRNALeu
mutant enzyme R286E, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
Escherichia coli
6.1.1.4
0.0025
-
Ile-tRNALeu
mutant enzyme E184R, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
Escherichia coli
6.1.1.4
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) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
6.1.1.4
ATP + L-leucine + tRNALeu
Escherichia coli
-
AMP + diphosphate + L-leucyl-tRNALeu
-
-
?
6.1.1.4
additional information
Escherichia coli
the enzyme has evolved both tRNA-dependent pre- and post-transfer editing capabilities to ensure catalytic specificity
?
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
6.1.1.4
Ni-NTA column chromatography
Escherichia coli
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
6.1.1.4
AMP + diphosphate + Ile-tRNALeu
-
726909
Escherichia coli
ATP + L-isoleucine + tRNALeu
-
-
-
?
6.1.1.4
ATP + L-leucine + tRNALeu
-
726909
Escherichia coli
AMP + diphosphate + L-leucyl-tRNALeu
-
-
-
?
6.1.1.4
additional information
the enzyme has evolved both tRNA-dependent pre- and post-transfer editing capabilities to ensure catalytic specificity
726909
Escherichia coli
?
-
-
-
?
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
6.1.1.4
7
-
Ile-tRNALeu
wild type enzyme, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
Escherichia coli
6.1.1.4
7.4
-
Ile-tRNALeu
mutant enzyme R286E, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
Escherichia coli
6.1.1.4
7.6
-
Ile-tRNALeu
mutant enzyme R185E, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
Escherichia coli
6.1.1.4
8.8
-
Ile-tRNALeu
mutant enzyme E184R, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
Escherichia coli
6.1.1.4
73.7
-
L-leucine
wild type enzyme, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
Escherichia coli
KCat/KM [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
6.1.1.4
1890
-
L-leucine
wild type enzyme, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
Escherichia coli
6.1.1.4
3100
-
Ile-tRNALeu
mutant enzyme R286E, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
Escherichia coli
6.1.1.4
3300
-
Ile-tRNALeu
wild type enzyme, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
Escherichia coli
6.1.1.4
3500
-
Ile-tRNALeu
mutant enzyme E184R, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
Escherichia coli
6.1.1.4
3800
-
Ile-tRNALeu
mutant enzyme R185E, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
Escherichia coli
KCat/KM [mM/s] (protein specific)
EC Number
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
6.1.1.4
1890
-
L-leucine
wild type enzyme, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
Escherichia coli
6.1.1.4
3100
-
Ile-tRNALeu
mutant enzyme R286E, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
Escherichia coli
6.1.1.4
3300
-
Ile-tRNALeu
wild type enzyme, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
Escherichia coli
6.1.1.4
3500
-
Ile-tRNALeu
mutant enzyme E184R, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
Escherichia coli
6.1.1.4
3800
-
Ile-tRNALeu
mutant enzyme R185E, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
Escherichia coli