BRENDA - Enzyme Database

Escherichia coli glutamyl-tRNA reductase. Trapping the thioester intermediate

Schauer, S.; Chaturvedi, S.; Randau, L.; Moser, J.; Kitabatake, M.; Lorenz, S.; Verkamp, E.; Schubert, W.D.; Nakayashiki, T.; Murai, M.; Wall, K.; Thomann, H.U.; Heinz, D.W.; Inokuchi, H.; Soll, D.; Jahn, D.; J. Biol. Chem. 277, 48657-48663 (2002)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
1.2.1.70
overexpression in Escherichia coli
Escherichia coli
Engineering
EC Number
Amino acid exchange
Commentary
Organism
1.2.1.70
C170S
mutant enzyme with esterase activity 95% of the wild-type activity and reductase activity with 90% of the wild-type activity
Escherichia coli
1.2.1.70
C50S
mutant enzyme with no esterase and reductase activity
Escherichia coli
1.2.1.70
C74S
mutant enzyme with esterase activity 110% of the wild-type activity and reductase activity with 120% of the wild-type activity
Escherichia coli
1.2.1.70
E114K
mutant enzyme with no esterase and reductase activity
Escherichia coli
1.2.1.70
G106N
mutant enzyme with no esterase and reductase activity
Escherichia coli
1.2.1.70
G191D
mutant enzyme reveals esterase, 105% of the wild-type activity, but no reductase activity
Escherichia coli
1.2.1.70
G44C/S105N/A326T
mutant enzyme with no esterase and reductase activity
Escherichia coli
1.2.1.70
G7D
mutant enzyme with no esterase and reductase activity
Escherichia coli
1.2.1.70
R314C
mutant enzyme with no esterase and reductase activity
Escherichia coli
1.2.1.70
S145F
mutant enzyme with no esterase and reductase activity
Escherichia coli
1.2.1.70
S22L/S164F
mutant enzyme with no esterase and reductase activity
Escherichia coli
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
1.2.1.70
1,10-phenanthroline
5 mM, 25% inhibition
Escherichia coli
1.2.1.70
2,2'-dipyridyl
5 mM, 20% inhibition
Escherichia coli
1.2.1.70
5,5'-dithiobis(2-nitrobenzoic acid)
1 mM, 80% inhibition
Escherichia coli
1.2.1.70
Co2+
-
Escherichia coli
1.2.1.70
EDTA
10 mM, 55% inhibition
Escherichia coli
1.2.1.70
EGTA
10 mM, 35% inhibition
Escherichia coli
1.2.1.70
glutamycin
3 mM, 90% inhibition
Escherichia coli
1.2.1.70
iodoacetamide
0.1 mM, complete inhibition
Escherichia coli
1.2.1.70
Ni2+
-
Escherichia coli
1.2.1.70
PtCl4
1 mM, 90% inhibition
Escherichia coli
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.2.1.70
0.024
-
L-glutamyl-tRNAGlu
pH 8.1, 37°C
Escherichia coli
1.2.1.70
0.039
-
NADPH
pH 8.1, 37°C
Escherichia coli
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
1.2.1.70
Ca2+
restores activity after treatment with chelating agents
Escherichia coli
1.2.1.70
Mg2+
stimulates, restores activity after treatment with chelating agents
Escherichia coli
1.2.1.70
Mn2+
restores activity after treatment with chelating agents
Escherichia coli
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.2.1.70
48000
-
2 * 48000, homodimer with an extended structure, SDS-PAGE
Escherichia coli
1.2.1.70
48448
-
2 * 48448, homodimer with an extended structure, electrospray ionization mass spectrometry
Escherichia coli
1.2.1.70
180000
-
gel filtration
Escherichia coli
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.2.1.70
Escherichia coli
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
1.2.1.70
-
Escherichia coli
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.2.1.70
L-glutamyl-tRNAGlu + NADPH + H+
in presence of NADPH, the end product, glutamate-1-semialdehyde is formed. In the absence of NADPH, Escherichia coli GluTR exhibits substrate esterase activity
656104
Escherichia coli
L-glutamate 1-semialdehyde + NADP+ + tRNAGlu
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
1.2.1.70
dimer
2 * 48000, homodimer with an extended structure, SDS-PAGE; 2 * 48448, homodimer with an extended structure, electrospray ionization mass spectrometry
Escherichia coli
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.2.1.70
0.13
-
L-glutamyl-tRNAGlu
pH 8.1, 37°C
Escherichia coli
1.2.1.70
0.15
-
NADPH
pH 8.1, 37°C
Escherichia coli
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.2.1.70
NADPH
-
Escherichia coli
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.2.1.70
overexpression in Escherichia coli
Escherichia coli
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.2.1.70
NADPH
-
Escherichia coli
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
1.2.1.70
C170S
mutant enzyme with esterase activity 95% of the wild-type activity and reductase activity with 90% of the wild-type activity
Escherichia coli
1.2.1.70
C50S
mutant enzyme with no esterase and reductase activity
Escherichia coli
1.2.1.70
C74S
mutant enzyme with esterase activity 110% of the wild-type activity and reductase activity with 120% of the wild-type activity
Escherichia coli
1.2.1.70
E114K
mutant enzyme with no esterase and reductase activity
Escherichia coli
1.2.1.70
G106N
mutant enzyme with no esterase and reductase activity
Escherichia coli
1.2.1.70
G191D
mutant enzyme reveals esterase, 105% of the wild-type activity, but no reductase activity
Escherichia coli
1.2.1.70
G44C/S105N/A326T
mutant enzyme with no esterase and reductase activity
Escherichia coli
1.2.1.70
G7D
mutant enzyme with no esterase and reductase activity
Escherichia coli
1.2.1.70
R314C
mutant enzyme with no esterase and reductase activity
Escherichia coli
1.2.1.70
S145F
mutant enzyme with no esterase and reductase activity
Escherichia coli
1.2.1.70
S22L/S164F
mutant enzyme with no esterase and reductase activity
Escherichia coli
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
1.2.1.70
1,10-phenanthroline
5 mM, 25% inhibition
Escherichia coli
1.2.1.70
2,2'-dipyridyl
5 mM, 20% inhibition
Escherichia coli
1.2.1.70
5,5'-dithiobis(2-nitrobenzoic acid)
1 mM, 80% inhibition
Escherichia coli
1.2.1.70
Co2+
-
Escherichia coli
1.2.1.70
EDTA
10 mM, 55% inhibition
Escherichia coli
1.2.1.70
EGTA
10 mM, 35% inhibition
Escherichia coli
1.2.1.70
glutamycin
3 mM, 90% inhibition
Escherichia coli
1.2.1.70
iodoacetamide
0.1 mM, complete inhibition
Escherichia coli
1.2.1.70
Ni2+
-
Escherichia coli
1.2.1.70
PtCl4
1 mM, 90% inhibition
Escherichia coli
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.2.1.70
0.024
-
L-glutamyl-tRNAGlu
pH 8.1, 37°C
Escherichia coli
1.2.1.70
0.039
-
NADPH
pH 8.1, 37°C
Escherichia coli
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
1.2.1.70
Ca2+
restores activity after treatment with chelating agents
Escherichia coli
1.2.1.70
Mg2+
stimulates, restores activity after treatment with chelating agents
Escherichia coli
1.2.1.70
Mn2+
restores activity after treatment with chelating agents
Escherichia coli
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.2.1.70
48000
-
2 * 48000, homodimer with an extended structure, SDS-PAGE
Escherichia coli
1.2.1.70
48448
-
2 * 48448, homodimer with an extended structure, electrospray ionization mass spectrometry
Escherichia coli
1.2.1.70
180000
-
gel filtration
Escherichia coli
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.2.1.70
-
Escherichia coli
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.2.1.70
L-glutamyl-tRNAGlu + NADPH + H+
in presence of NADPH, the end product, glutamate-1-semialdehyde is formed. In the absence of NADPH, Escherichia coli GluTR exhibits substrate esterase activity
656104
Escherichia coli
L-glutamate 1-semialdehyde + NADP+ + tRNAGlu
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.2.1.70
dimer
2 * 48000, homodimer with an extended structure, SDS-PAGE; 2 * 48448, homodimer with an extended structure, electrospray ionization mass spectrometry
Escherichia coli
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.2.1.70
0.13
-
L-glutamyl-tRNAGlu
pH 8.1, 37°C
Escherichia coli
1.2.1.70
0.15
-
NADPH
pH 8.1, 37°C
Escherichia coli