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Literature summary for 1.4.1.13 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
Search for more literature for 1.4.1.13

General Stability

General Stability Organism
2-mercaptoethanol, stabilizes Escherichia coli
alpha-ketoglutarate stabilizes Escherichia coli
alpha-ketoglutarate, L-glutamine, L-glutamate and NADP when present together partial protect against enzyme inactivation Escherichia coli
incubation with L-glutamine or L-glutamate results in marked time- and temperature-dependent loss in activity Escherichia coli
NADP causes relatively small loss in activity compared with control values Escherichia coli

Inhibitors

Inhibitors Comment Organism Structure
ADP 15% inhibition at 50 mM Escherichia coli
ATP 19% inhibition at 25 mM and 43% inhibition at 50 mM Escherichia coli
BaCl2 61% inhibition at 50 mM Escherichia coli
CaCl2 74% inhibition at 50 mM Escherichia coli
CdCl2 produces more than 60% inhibition at 5 mM Escherichia coli
cis-aconitate 22% inhibition at 50 mM Escherichia coli
CoCl2 produces more than 60% inhibition at 5 mM Escherichia coli
CTP 9% inhibition at 25 mM and 21% inhibition at 50 mM Escherichia coli
D-glutamate 50% inhibition at concentrations below 7 mM Escherichia coli
D-methionine 19% inhibition at 50 mM Escherichia coli
fumarate 21% inhibition at 50 mM Escherichia coli
glycine significant inhibitor Escherichia coli
GTP 16% inhibition at 25 mM and 43% inhibition at 50 mM Escherichia coli
iodoacetamide more than 50% loss in activity in 60 min at 0.5 mM Escherichia coli
isocitrate 35% inhibition at 50 mM Escherichia coli
ITP 17% inhibition at 25 mM and 29% inhibition at 50 mM Escherichia coli
KBr 43% inhibition at 100 mM Escherichia coli
KCl 32% inhibition at 100 mM Escherichia coli
KNO3 47% inhibition at 100 mM Escherichia coli
L-alanine significant inhibitor Escherichia coli
L-asparagine significant inhibitor Escherichia coli
L-aspartate 50% inhibition at concentrations below 7 mM Escherichia coli
L-cysteine significant inhibitor Escherichia coli
L-glutamate 50% inhibition at concentrations between 17 and 35 mM Escherichia coli
L-histidine significant inhibitor Escherichia coli
L-homoserine significant inhibitor Escherichia coli
L-malate 19% inhibition at 50 mM Escherichia coli
L-methionine 50% inhibition at concentrations below 7 mM Escherichia coli
L-serine significant inhibitor Escherichia coli
MgCl2 70% inhibition at 50 mM Escherichia coli
MnCl2 82% inhibition at 50 mM Escherichia coli
NADP+ 50% inhibition at concentrations below 7 mM Escherichia coli
NiCl2 produces more than 60% inhibition at 5 mM Escherichia coli
oxaloacetate
-
 Escherichia coli
p-chloromercuriphenylsulfonate more than 50% loss in activity in 60 min at 0.05 mM Escherichia coli
pyruvate 20% inhibition at 50 mM Escherichia coli
succinate 16% inhibition at 50 mM Escherichia coli
UTP 17% inhibition at 25 mM and 33% inhibition at 50 mM Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0073
-
 alpha-ketoglutarate
-
 Escherichia coli
0.0077
-
 NADPH
-
 Escherichia coli
0.25
-
 L-glutamine
-
 Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
Iron iron-sulfur protein Escherichia coli
Iron contains 38.4 mol of iron Escherichia coli

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
53000
-
 4 * 53000 + 4 * 135000, SDS-PAGE, urea polyacrylamide gel electrophoresis Escherichia coli
135000
-
 4 * 53000 + 4 * 135000, SDS-PAGE, urea polyacrylamide gel electrophoresis Escherichia coli
800000
-
 sedimentation equilibrium, gel filtration Escherichia coli

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-

Purification (Commentary)

Purification (Comment) Organism
using streptomycin sulfate treatment, ammonium sulfate fractionation, heat treatment, agarose gel filtration and DEAE-cellulose column chromatography Escherichia coli

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
26.2
-
-
 Escherichia coli

Storage Stability

Storage Stability Organism
-80°C, 20 mM potassium-HEPES, pH 7.2, 2 mM alpha-ketoglutarate, 1 mM EDTA, stable for at least 6 months Escherichia coli
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)

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

Subunits Comment Organism
octamer 4 * 53000 + 4 * 135000, SDS-PAGE, urea polyacrylamide gel electrophoresis Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.6
-
-
 Escherichia coli

pH Range

pH Minimum pH Maximum Comment Organism
6.4 9 at pH 6.4 and 9.0 about 50% of activity maximum Escherichia coli
6.5 8.7 at pH 6.5 and 8.7 about 50% of activity maximum Escherichia coli

Cofactor

Cofactor Comment Organism Structure
flavin flavoenzyme Escherichia coli
flavin contains FAD and FMN Escherichia coli
flavin ratio FAD: FMN is 1.2 Escherichia coli
flavin enzyme contains 7.8 mol of flavin Escherichia coli
additional information purified enzyme contains 30.4 mol of labile sulfide per 800,000 g of protein Escherichia coli
NADH no activity with NADH Escherichia coli
NADPH
-
 Escherichia coli