Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 1.4.1.4 extracted from

  • Zocher, K.; Fritz-Wolf, K.; Kehr, S.; Fischer, M.; Rahlfs, S.; Becker, K.
    Biochemical and structural characterization of Plasmodium falciparum glutamate dehydrogenase 2 (2012), Mol. Biochem. Parasitol., 183, 52-62.
    View publication on PubMed
Search for more literature for 1.4.1.4

Cloned(Commentary)

Cloned (Comment) Organism
gene GDH1, DNA and amino acid sequence determination and analysis, overexpression as His6-tagged protein in Escherichia coli strain KRX Plasmodium falciparum
gene GDH1, expression as His6-tagged enzyme in Escherichia coli strain M15, method optimization Plasmodium falciparum

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant detagged GDH2, 0.0025 ml of 16 mg/ml protein in 100 mM Tris, 500 mM NaCl, pH 7.8, mixed with 0.0025 ml of reservoir solution containing 0.2 M magnesium chloride hexahydrate, 0.1 M Tris/HCl pH 8.5, 30% PEG 4000 including 0.01 M spermine tetra HCl droplet concentration and 10 mM glutamate, X-ray diffraction structure determination and analysis at 3.1 A resolution Plasmodium falciparum

Inhibitors

Inhibitors Comment Organism Structure
bithionol
-
 Plasmodium falciparum
GW5074
-
 Plasmodium falciparum
Hexachlorophene 15% inhibition at 0.016 mM; 80% inhibition at 0.016 mM Plasmodium falciparum
additional information inhibitors hexachlorophene, GW5074, and bithionol are more effective on PfGDH2 than on PfGDH1; inhibitors hexachlorophene, GW5074, and bithionol are more effective on PfGDH2 than on PfGDH1 Plasmodium falciparum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.007
-
 NADPH pH 7.0, 25°C, recombinant enzyme Plasmodium falciparum
0.012
-
 NADPH pH 7.0, 25°C, recombinant enzyme Plasmodium falciparum
0.019
-
 NADP+ pH 8.0, 25°C, recombinant enzyme Plasmodium falciparum
0.02
-
 NADP+ pH 8.0, 25°C, recombinant enzyme Plasmodium falciparum
0.2
-
 2-oxoglutarate pH 7.0, 25°C, recombinant enzyme Plasmodium falciparum
0.365
-
 2-oxoglutarate pH 7.0, 25°C, recombinant enzyme Plasmodium falciparum
0.61
-
 L-glutamate pH 8.0, 25°C, recombinant enzyme Plasmodium falciparum
1.05
-
 L-glutamate pH 8.0, 25°C, recombinant enzyme Plasmodium falciparum

Localization

Localization Comment Organism GeneOntology No. Textmining
apicoplast GDH2 co-localizes with the acyl carrier protein Plasmodium falciparum 20011
-
cytosol
-
 Plasmodium falciparum 5829
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
52546
-
 6 * 52546, GDH1, sequence calculation Plasmodium falciparum
54024
-
 6 * 54024, GDH2, sequence calculation Plasmodium falciparum
300000
-
 recombinant enzyme, gel filtration Plasmodium falciparum

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2-oxoglutarate + NH3 + NADPH + H+ Plasmodium falciparum
-
 L-glutamate + H2O + NADP+
-
 r
L-glutamate + H2O + NADP+ Plasmodium falciparum
-
 2-oxoglutarate + NH3 + NADPH + H+
-
 r
additional information Plasmodium falciparum Like PfGDH1, PfGDH2 is an NADP(H)-dependent enzyme with a specific activity comparable to PfGDH1 but with slightly higher Km values for its substrates ?
-
 ?
additional information Plasmodium falciparum PfGDH1, like PfGDH2, is an NADP(H)-dependent enzyme with a specific activity comparable to PfGDH2 but with slightly lower Km values for its substrates ?
-
 ?

Organism

Organism UniProt Comment Textmining
Plasmodium falciparum Q8ILF7 GDH2
-
Plasmodium falciparum Q8ILT0 GDH1
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His6-tagged GDH2 from Escherichia coli strain KRX by nickel affinity chromatography and gel filtration Plasmodium falciparum

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
4
-
 deamination, pH 8.0, 25°C, recombinant enzyme Plasmodium falciparum
9
-
 deamination, pH 8.0, 25°C, recombinant enzyme Plasmodium falciparum
28
-
 amination, pH 8.0, 25°C, recombinant enzyme Plasmodium falciparum
61
-
 amination, pH 8.0, 25°C, recombinant enzyme Plasmodium falciparum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-oxoglutarate + NH3 + NADPH + H+
-
 Plasmodium falciparum L-glutamate + H2O + NADP+
-
 r
2-oxoglutarate + NH3 + NADPH + H+ referred reaction Plasmodium falciparum L-glutamate + H2O + NADP+
-
 r
L-glutamate + H2O + NADP+
-
 Plasmodium falciparum 2-oxoglutarate + NH3 + NADPH + H+
-
 r
additional information Like PfGDH1, PfGDH2 is an NADP(H)-dependent enzyme with a specific activity comparable to PfGDH1 but with slightly higher Km values for its substrates Plasmodium falciparum ?
-
 ?
additional information PfGDH1, like PfGDH2, is an NADP(H)-dependent enzyme with a specific activity comparable to PfGDH2 but with slightly lower Km values for its substrates Plasmodium falciparum ?
-
 ?

Subunits

Subunits Comment Organism
hexamer 6 * 52546, GDH1, sequence calculation Plasmodium falciparum
hexamer 6 * 54024, GDH2, sequence calculation Plasmodium falciparum
More the subunit interactions are mediated by networks of salt bridges and hydrogen bonds, monome-monomer interaction, overview Plasmodium falciparum
More the three-dimensional structure of hexameric PfGDH2 is solved to 3.1 A resolution, overview. The subunit interactions are mainly assisted by hydrogen bonds and hydrophobic interactions, monome-monomer interaction, overview Plasmodium falciparum

Synonyms

Synonyms Comment Organism
GDH1
-
 Plasmodium falciparum
GDH2
-
 Plasmodium falciparum
glutamate dehydrogenase 1
-
 Plasmodium falciparum
glutamate dehydrogenase 2
-
 Plasmodium falciparum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
 assay at Plasmodium falciparum
25
-
 amination, assay at Plasmodium falciparum

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
80
-
 purified enzyme, 10 min, stable Plasmodium falciparum

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.367
-
 L-glutamate pH 8.0, 25°C, recombinant enzyme Plasmodium falciparum
25
-
 2-oxoglutarate pH 7.0, 25°C, recombinant enzyme Plasmodium falciparum
53.33
-
 2-oxoglutarate pH 7.0, 25°C, recombinant enzyme Plasmodium falciparum
78.33
-
 L-glutamate pH 8.0, 25°C, recombinant enzyme Plasmodium falciparum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
 amination, assay at Plasmodium falciparum
8
-
 deamination, assay at Plasmodium falciparum

Cofactor

Cofactor Comment Organism Structure
additional information no activity with NAD+ Plasmodium falciparum
NADP+
-
 Plasmodium falciparum
NADPH
-
 Plasmodium falciparum

pI Value

Organism Comment pI Value Maximum pI Value
Plasmodium falciparum sequence calculation
-
 7.48
Plasmodium falciparum sequence calculation
-
 9.09

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
0.0252
-
 pH 8.0, 25°C, recombinant enzyme Plasmodium falciparum bithionol
0.12
-
 pH 8.0, 25°C, recombinant enzyme Plasmodium falciparum bithionol
0.139
-
 pH 8.0, 25°C, recombinant enzyme Plasmodium falciparum GW5074
0.15
-
 pH 8.0, 25°C, recombinant enzyme Plasmodium falciparum GW5074

General Information

General Information Comment Organism
additional information the parasitic enzyme does not contain the antenna domain, responsible for allosteric regulation in the mammalian enzymes Plasmodium falciparum
additional information the three-dimensional structure of hexameric PfGDH2 is solved to 3.1 A resolution, overview. The parasitic enzyme does not contain the antenna domain, responsible for allosteric regulation in the mammalian enzymes Plasmodium falciparum

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.75
-
 L-glutamate pH 8.0, 25°C, recombinant enzyme Plasmodium falciparum
6
-
 L-glutamate pH 8.0, 25°C, recombinant enzyme Plasmodium falciparum
14.67
-
 2-oxoglutarate pH 7.0, 25°C, recombinant enzyme Plasmodium falciparum
1250
-
 2-oxoglutarate pH 7.0, 25°C, recombinant enzyme Plasmodium falciparum