BRENDA - Enzyme Database

Glutamate synthase from Escherichia coli. An iron-sulfide flavoprotein

Miller, R.E.; Stadtman, E.R.; J. Biol. Chem. 247, 7407-7419 (1972)

Data extracted from this reference:

General Stability
EC Number
General Stability
Organism
1.4.1.13
2-mercaptoethanol, stabilizes
Escherichia coli
1.4.1.13
alpha-ketoglutarate stabilizes
Escherichia coli
1.4.1.13
alpha-ketoglutarate, L-glutamine, L-glutamate and NADP when present together partial protect against enzyme inactivation
Escherichia coli
1.4.1.13
incubation with L-glutamine or L-glutamate results in marked time- and temperature-dependent loss in activity
Escherichia coli
1.4.1.13
NADP causes relatively small loss in activity compared with control values
Escherichia coli
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
1.4.1.13
ADP
15% inhibition at 50 mM
Escherichia coli
1.4.1.13
ATP
19% inhibition at 25 mM and 43% inhibition at 50 mM
Escherichia coli
1.4.1.13
BaCl2
61% inhibition at 50 mM
Escherichia coli
1.4.1.13
CaCl2
74% inhibition at 50 mM
Escherichia coli
1.4.1.13
CdCl2
produces more than 60% inhibition at 5 mM
Escherichia coli
1.4.1.13
cis-aconitate
22% inhibition at 50 mM
Escherichia coli
1.4.1.13
CoCl2
produces more than 60% inhibition at 5 mM
Escherichia coli
1.4.1.13
CTP
9% inhibition at 25 mM and 21% inhibition at 50 mM
Escherichia coli
1.4.1.13
D-glutamate
50% inhibition at concentrations below 7 mM
Escherichia coli
1.4.1.13
D-methionine
19% inhibition at 50 mM
Escherichia coli
1.4.1.13
fumarate
21% inhibition at 50 mM
Escherichia coli
1.4.1.13
glycine
significant inhibitor
Escherichia coli
1.4.1.13
GTP
16% inhibition at 25 mM and 43% inhibition at 50 mM
Escherichia coli
1.4.1.13
iodoacetamide
more than 50% loss in activity in 60 min at 0.5 mM
Escherichia coli
1.4.1.13
isocitrate
35% inhibition at 50 mM
Escherichia coli
1.4.1.13
ITP
17% inhibition at 25 mM and 29% inhibition at 50 mM
Escherichia coli
1.4.1.13
KBr
43% inhibition at 100 mM
Escherichia coli
1.4.1.13
KCl
32% inhibition at 100 mM
Escherichia coli
1.4.1.13
KNO3
47% inhibition at 100 mM
Escherichia coli
1.4.1.13
L-alanine
significant inhibitor
Escherichia coli
1.4.1.13
L-asparagine
significant inhibitor
Escherichia coli
1.4.1.13
L-aspartate
50% inhibition at concentrations below 7 mM
Escherichia coli
1.4.1.13
L-cysteine
significant inhibitor
Escherichia coli
1.4.1.13
L-glutamate
50% inhibition at concentrations between 17 and 35 mM
Escherichia coli
1.4.1.13
L-histidine
significant inhibitor
Escherichia coli
1.4.1.13
L-homoserine
significant inhibitor
Escherichia coli
1.4.1.13
L-malate
19% inhibition at 50 mM
Escherichia coli
1.4.1.13
L-methionine
50% inhibition at concentrations below 7 mM
Escherichia coli
1.4.1.13
L-serine
significant inhibitor
Escherichia coli
1.4.1.13
MgCl2
70% inhibition at 50 mM
Escherichia coli
1.4.1.13
MnCl2
82% inhibition at 50 mM
Escherichia coli
1.4.1.13
NADP+
50% inhibition at concentrations below 7 mM
Escherichia coli
1.4.1.13
NiCl2
produces more than 60% inhibition at 5 mM
Escherichia coli
1.4.1.13
oxaloacetate
-
Escherichia coli
1.4.1.13
p-chloromercuriphenylsulfonate
more than 50% loss in activity in 60 min at 0.05 mM
Escherichia coli
1.4.1.13
pyruvate
20% inhibition at 50 mM
Escherichia coli
1.4.1.13
succinate
16% inhibition at 50 mM
Escherichia coli
1.4.1.13
UTP
17% inhibition at 25 mM and 33% inhibition at 50 mM
Escherichia coli
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.4.1.13
0.0073
-
alpha-ketoglutarate
-
Escherichia coli
1.4.1.13
0.0077
-
NADPH
-
Escherichia coli
1.4.1.13
0.25
-
L-glutamine
-
Escherichia coli
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
1.4.1.13
Iron
contains 38.4 mol of iron; iron-sulfur protein
Escherichia coli
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.4.1.13
53000
-
4 * 53000 + 4 * 135000, SDS-PAGE, urea polyacrylamide gel electrophoresis
Escherichia coli
1.4.1.13
135000
-
4 * 53000 + 4 * 135000, SDS-PAGE, urea polyacrylamide gel electrophoresis
Escherichia coli
1.4.1.13
800000
-
sedimentation equilibrium, gel filtration
Escherichia coli
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.4.1.13
Escherichia coli
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
1.4.1.13
using streptomycin sulfate treatment, ammonium sulfate fractionation, heat treatment, agarose gel filtration and DEAE-cellulose column chromatography
Escherichia coli
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.4.1.13
26.2
-
-
Escherichia coli
Storage Stability
EC Number
Storage Stability
Organism
1.4.1.13
-80°C, 20 mM potassium-HEPES, pH 7.2, 2 mM alpha-ketoglutarate, 1 mM EDTA, stable for at least 6 months
Escherichia coli
1.4.1.13
4°C, 20 mM potassium-HEPES, pH 7.2, 2 mM alpha-ketoglutarate, 1 mM EDTA, 5 mM 2-mercaptoethanol, 20% loss of activity, after 1 month
Escherichia coli
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.4.1.13
L-glutamine + 2-oxoglutarate + NADPH + H+
highly specific
391438
Escherichia coli
L-glutamate + NADP+
-
391438
Escherichia coli
ir
1.4.1.13
L-glutamine + 2-oxoglutarate + NADPH + H+
L-glutamine, 2-oxoglutarate and NADPH are all required for catalytic activity
391438
Escherichia coli
L-glutamate + NADP+
-
391438
Escherichia coli
ir
1.4.1.13
additional information
NH4Cl, L-asparagine, D-glutamine, or alkylated glutamine analogues do not substitute for L-glutamine, pyruvate or oxalacetate do not substitute for alpha-ketoglutarate
391438
Escherichia coli
?
-
-
-
-
Subunits
EC Number
Subunits
Commentary
Organism
1.4.1.13
octamer
4 * 53000 + 4 * 135000, SDS-PAGE, urea polyacrylamide gel electrophoresis
Escherichia coli
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.4.1.13
7.6
-
-
Escherichia coli
pH Range
EC Number
pH Minimum
pH Maximum
Commentary
Organism
1.4.1.13
6.4
9
at pH 6.4 and 9.0 about 50% of activity maximum
Escherichia coli
1.4.1.13
6.5
8.7
at pH 6.5 and 8.7 about 50% of activity maximum
Escherichia coli
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.4.1.13
flavin
contains FAD and FMN; enzyme contains 7.8 mol of flavin; flavoenzyme; ratio FAD: FMN is 1.2
Escherichia coli
1.4.1.13
additional information
purified enzyme contains 30.4 mol of labile sulfide per 800,000 g of protein
Escherichia coli
1.4.1.13
NADH
no activity with NADH
Escherichia coli
1.4.1.13
NADPH
-
Escherichia coli
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.4.1.13
flavin
contains FAD and FMN; enzyme contains 7.8 mol of flavin; flavoenzyme; ratio FAD: FMN is 1.2
Escherichia coli
1.4.1.13
additional information
purified enzyme contains 30.4 mol of labile sulfide per 800,000 g of protein
Escherichia coli
1.4.1.13
NADH
no activity with NADH
Escherichia coli
1.4.1.13
NADPH
-
Escherichia coli
General Stability (protein specific)
EC Number
General Stability
Organism
1.4.1.13
2-mercaptoethanol, stabilizes
Escherichia coli
1.4.1.13
alpha-ketoglutarate stabilizes
Escherichia coli
1.4.1.13
alpha-ketoglutarate, L-glutamine, L-glutamate and NADP when present together partial protect against enzyme inactivation
Escherichia coli
1.4.1.13
incubation with L-glutamine or L-glutamate results in marked time- and temperature-dependent loss in activity
Escherichia coli
1.4.1.13
NADP causes relatively small loss in activity compared with control values
Escherichia coli
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
1.4.1.13
ADP
15% inhibition at 50 mM
Escherichia coli
1.4.1.13
ATP
19% inhibition at 25 mM and 43% inhibition at 50 mM
Escherichia coli
1.4.1.13
BaCl2
61% inhibition at 50 mM
Escherichia coli
1.4.1.13
CaCl2
74% inhibition at 50 mM
Escherichia coli
1.4.1.13
CdCl2
produces more than 60% inhibition at 5 mM
Escherichia coli
1.4.1.13
cis-aconitate
22% inhibition at 50 mM
Escherichia coli
1.4.1.13
CoCl2
produces more than 60% inhibition at 5 mM
Escherichia coli
1.4.1.13
CTP
9% inhibition at 25 mM and 21% inhibition at 50 mM
Escherichia coli
1.4.1.13
D-glutamate
50% inhibition at concentrations below 7 mM
Escherichia coli
1.4.1.13
D-methionine
19% inhibition at 50 mM
Escherichia coli
1.4.1.13
fumarate
21% inhibition at 50 mM
Escherichia coli
1.4.1.13
glycine
significant inhibitor
Escherichia coli
1.4.1.13
GTP
16% inhibition at 25 mM and 43% inhibition at 50 mM
Escherichia coli
1.4.1.13
iodoacetamide
more than 50% loss in activity in 60 min at 0.5 mM
Escherichia coli
1.4.1.13
isocitrate
35% inhibition at 50 mM
Escherichia coli
1.4.1.13
ITP
17% inhibition at 25 mM and 29% inhibition at 50 mM
Escherichia coli
1.4.1.13
KBr
43% inhibition at 100 mM
Escherichia coli
1.4.1.13
KCl
32% inhibition at 100 mM
Escherichia coli
1.4.1.13
KNO3
47% inhibition at 100 mM
Escherichia coli
1.4.1.13
L-alanine
significant inhibitor
Escherichia coli
1.4.1.13
L-asparagine
significant inhibitor
Escherichia coli
1.4.1.13
L-aspartate
50% inhibition at concentrations below 7 mM
Escherichia coli
1.4.1.13
L-cysteine
significant inhibitor
Escherichia coli
1.4.1.13
L-glutamate
50% inhibition at concentrations between 17 and 35 mM
Escherichia coli
1.4.1.13
L-histidine
significant inhibitor
Escherichia coli
1.4.1.13
L-homoserine
significant inhibitor
Escherichia coli
1.4.1.13
L-malate
19% inhibition at 50 mM
Escherichia coli
1.4.1.13
L-methionine
50% inhibition at concentrations below 7 mM
Escherichia coli
1.4.1.13
L-serine
significant inhibitor
Escherichia coli
1.4.1.13
MgCl2
70% inhibition at 50 mM
Escherichia coli
1.4.1.13
MnCl2
82% inhibition at 50 mM
Escherichia coli
1.4.1.13
NADP+
50% inhibition at concentrations below 7 mM
Escherichia coli
1.4.1.13
NiCl2
produces more than 60% inhibition at 5 mM
Escherichia coli
1.4.1.13
oxaloacetate
-
Escherichia coli
1.4.1.13
p-chloromercuriphenylsulfonate
more than 50% loss in activity in 60 min at 0.05 mM
Escherichia coli
1.4.1.13
pyruvate
20% inhibition at 50 mM
Escherichia coli
1.4.1.13
succinate
16% inhibition at 50 mM
Escherichia coli
1.4.1.13
UTP
17% inhibition at 25 mM and 33% inhibition at 50 mM
Escherichia coli
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.4.1.13
0.0073
-
alpha-ketoglutarate
-
Escherichia coli
1.4.1.13
0.0077
-
NADPH
-
Escherichia coli
1.4.1.13
0.25
-
L-glutamine
-
Escherichia coli
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
1.4.1.13
Iron
contains 38.4 mol of iron; iron-sulfur protein
Escherichia coli
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.4.1.13
53000
-
4 * 53000 + 4 * 135000, SDS-PAGE, urea polyacrylamide gel electrophoresis
Escherichia coli
1.4.1.13
135000
-
4 * 53000 + 4 * 135000, SDS-PAGE, urea polyacrylamide gel electrophoresis
Escherichia coli
1.4.1.13
800000
-
sedimentation equilibrium, gel filtration
Escherichia coli
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.4.1.13
using streptomycin sulfate treatment, ammonium sulfate fractionation, heat treatment, agarose gel filtration and DEAE-cellulose column chromatography
Escherichia coli
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.4.1.13
26.2
-
-
Escherichia coli
Storage Stability (protein specific)
EC Number
Storage Stability
Organism
1.4.1.13
-80°C, 20 mM potassium-HEPES, pH 7.2, 2 mM alpha-ketoglutarate, 1 mM EDTA, stable for at least 6 months
Escherichia coli
1.4.1.13
4°C, 20 mM potassium-HEPES, pH 7.2, 2 mM alpha-ketoglutarate, 1 mM EDTA, 5 mM 2-mercaptoethanol, 20% loss of activity, after 1 month
Escherichia coli
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.4.1.13
L-glutamine + 2-oxoglutarate + NADPH + H+
highly specific
391438
Escherichia coli
L-glutamate + NADP+
-
391438
Escherichia coli
ir
1.4.1.13
L-glutamine + 2-oxoglutarate + NADPH + H+
L-glutamine, 2-oxoglutarate and NADPH are all required for catalytic activity
391438
Escherichia coli
L-glutamate + NADP+
-
391438
Escherichia coli
ir
1.4.1.13
additional information
NH4Cl, L-asparagine, D-glutamine, or alkylated glutamine analogues do not substitute for L-glutamine, pyruvate or oxalacetate do not substitute for alpha-ketoglutarate
391438
Escherichia coli
?
-
-
-
-
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.4.1.13
octamer
4 * 53000 + 4 * 135000, SDS-PAGE, urea polyacrylamide gel electrophoresis
Escherichia coli
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.4.1.13
7.6
-
-
Escherichia coli
pH Range (protein specific)
EC Number
pH Minimum
pH Maximum
Commentary
Organism
1.4.1.13
6.4
9
at pH 6.4 and 9.0 about 50% of activity maximum
Escherichia coli
1.4.1.13
6.5
8.7
at pH 6.5 and 8.7 about 50% of activity maximum
Escherichia coli