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Literature summary extracted from

  • Miller, R.E.; Stadtman, E.R.
    Glutamate synthase from Escherichia coli. An iron-sulfide flavoprotein (1972), J. Biol. Chem., 247, 7407-7419.
    View publication on PubMed

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 Comment 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 Comment 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 Comment Organism Structure
1.4.1.13 Iron iron-sulfur protein Escherichia coli
1.4.1.13 Iron contains 38.4 mol of iron Escherichia coli

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment 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 UniProt Comment Textmining
1.4.1.13 Escherichia coli
-
-
-

Purification (Commentary)

EC Number Purification (Comment) 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] Comment 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 Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.4.1.13 L-glutamine + 2-oxoglutarate + NADPH + H+ highly specific Escherichia coli L-glutamate + NADP+
-
ir
1.4.1.13 L-glutamine + 2-oxoglutarate + NADPH + H+ L-glutamine, 2-oxoglutarate and NADPH are all required for catalytic activity Escherichia coli L-glutamate + NADP+
-
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 Escherichia coli ?
-
?

Subunits

EC Number Subunits Comment 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 Comment Organism
1.4.1.13 7.6
-
-
Escherichia coli

pH Range

EC Number pH Minimum pH Maximum Comment 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 Comment Organism Structure
1.4.1.13 flavin flavoenzyme Escherichia coli
1.4.1.13 flavin contains FAD and FMN Escherichia coli
1.4.1.13 flavin ratio FAD: FMN is 1.2 Escherichia coli
1.4.1.13 flavin enzyme contains 7.8 mol of flavin 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