BRENDA - Enzyme Database

Structural basis for anticodon recognition by discriminating glutamyl-tRNA synthetase

Sekine, S.; Nureki, O.; Shimada, A.; Vassylyev, D.G.; Yokoyama, S.; Nat. Struct. Biol. 8, 203-206 (2001)

Data extracted from this reference:

Crystallization (Commentary)
EC Number
Crystallization
Organism
6.1.1.17
purified recombinant enzyme in complex with tRNAGlu, hanging-drop vapour diffusion method, precipitant solution contains 37 mM Na-MOPS, pH 6.7, 22% PEG 1500, 37 mM ammonium sulfate, 1% 2-methyl-2,4-pentanediol, 10 mM MgCl2, 5 mM 2-mercaptoethanol, X-ray diffraction structure determination at 2.4 A resolution, and analysis
Thermus thermophilus
Engineering
EC Number
Amino acid exchange
Commentary
Organism
6.1.1.17
R358Q
site-directed mutagenesis, exchange of the Arg residue results in a mutant that no longer discriminates between tRNAGlu and tRNAGln anticodons YUC and YUG, respectively
Thermus thermophilus
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
6.1.1.17
0.0047
-
wild-type tRNAGlu
recombinant wild-type enzyme, 65°C
Thermus thermophilus
6.1.1.17
0.043
-
C36G mutant tRNAGlu
recombinant wild-type enzyme, 65°C
Thermus thermophilus
6.1.1.17
0.055
-
C36G mutant tRNAGlu
recombinant mutant R358Q, 65°C
Thermus thermophilus
6.1.1.17
0.085
-
wild-type tRNAGlu
recombinant mutant R358Q, 65°C
Thermus thermophilus
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
6.1.1.17
ATP + L-glutamate + tRNAGlu
Thermus thermophilus
-
AMP + diphosphate + L-glutamyl-tRNAGlu
-
Thermus thermophilus
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
6.1.1.17
Thermus thermophilus
P27000
discriminating enzyme form, purified recombinant enzyme from overexpressing Escherichia coli strain JM109(DE3)
-
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
6.1.1.17
ATP + L-glutamate + tRNAGlu
-
653317
Thermus thermophilus
AMP + diphosphate + L-glutamyl-tRNAGlu
-
653317
Thermus thermophilus
?
6.1.1.17
ATP + L-glutamate + tRNAGlu mutant C36G
mutant R358Q, low activity with the wild-type enzyme
653317
Thermus thermophilus
AMP + diphosphate + L-glutamyl-tRNAGlu mutant C36G
-
653317
Thermus thermophilus
?
6.1.1.17
ATP + L-glutamate + tRNAGlu wild-type
enzyme is specific for tRNAGlu
653317
Thermus thermophilus
AMP + diphosphate + L-glutamyl-tRNAGlu wild-type
-
653317
Thermus thermophilus
?
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
6.1.1.17
65
-
assay at
Thermus thermophilus
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
6.1.1.17
0.18
-
C36G mutant tRNAGlu
recombinant wild-type enzyme, 65°C
Thermus thermophilus
6.1.1.17
1.4
-
C36G mutant tRNAGlu
recombinant mutant enzyme R358Q, 65°C
Thermus thermophilus
6.1.1.17
1.5
-
wild-type tRNAGlu
recombinant mutant enzyme R358Q, 65°C
Thermus thermophilus
6.1.1.17
2.1
-
wild-type tRNAGlu
recombinant wild-type enzyme, 65°C
Thermus thermophilus
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
6.1.1.17
ATP
-
Thermus thermophilus
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
6.1.1.17
ATP
-
Thermus thermophilus
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
6.1.1.17
purified recombinant enzyme in complex with tRNAGlu, hanging-drop vapour diffusion method, precipitant solution contains 37 mM Na-MOPS, pH 6.7, 22% PEG 1500, 37 mM ammonium sulfate, 1% 2-methyl-2,4-pentanediol, 10 mM MgCl2, 5 mM 2-mercaptoethanol, X-ray diffraction structure determination at 2.4 A resolution, and analysis
Thermus thermophilus
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
6.1.1.17
R358Q
site-directed mutagenesis, exchange of the Arg residue results in a mutant that no longer discriminates between tRNAGlu and tRNAGln anticodons YUC and YUG, respectively
Thermus thermophilus
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
6.1.1.17
0.0047
-
wild-type tRNAGlu
recombinant wild-type enzyme, 65°C
Thermus thermophilus
6.1.1.17
0.043
-
C36G mutant tRNAGlu
recombinant wild-type enzyme, 65°C
Thermus thermophilus
6.1.1.17
0.055
-
C36G mutant tRNAGlu
recombinant mutant R358Q, 65°C
Thermus thermophilus
6.1.1.17
0.085
-
wild-type tRNAGlu
recombinant mutant R358Q, 65°C
Thermus thermophilus
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
6.1.1.17
ATP + L-glutamate + tRNAGlu
Thermus thermophilus
-
AMP + diphosphate + L-glutamyl-tRNAGlu
-
Thermus thermophilus
?
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
6.1.1.17
ATP + L-glutamate + tRNAGlu
-
653317
Thermus thermophilus
AMP + diphosphate + L-glutamyl-tRNAGlu
-
653317
Thermus thermophilus
?
6.1.1.17
ATP + L-glutamate + tRNAGlu mutant C36G
mutant R358Q, low activity with the wild-type enzyme
653317
Thermus thermophilus
AMP + diphosphate + L-glutamyl-tRNAGlu mutant C36G
-
653317
Thermus thermophilus
?
6.1.1.17
ATP + L-glutamate + tRNAGlu wild-type
enzyme is specific for tRNAGlu
653317
Thermus thermophilus
AMP + diphosphate + L-glutamyl-tRNAGlu wild-type
-
653317
Thermus thermophilus
?
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
6.1.1.17
65
-
assay at
Thermus thermophilus
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
6.1.1.17
0.18
-
C36G mutant tRNAGlu
recombinant wild-type enzyme, 65°C
Thermus thermophilus
6.1.1.17
1.4
-
C36G mutant tRNAGlu
recombinant mutant enzyme R358Q, 65°C
Thermus thermophilus
6.1.1.17
1.5
-
wild-type tRNAGlu
recombinant mutant enzyme R358Q, 65°C
Thermus thermophilus
6.1.1.17
2.1
-
wild-type tRNAGlu
recombinant wild-type enzyme, 65°C
Thermus thermophilus