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Literature summary for 1.4.1.4 extracted from

  • Hou, W.; Lu, S.; Zhao, H.; Yu, Y.; Xu, H.; Yu, B.; Su, L.; Lin, C.; Ruan, B.H.
    Propylselen inhibits cancer cell growth by targeting glutamate dehydrogenase at the NADP+ binding site (2019), Biochem. Biophys. Res. Commun., 509, 262-267 .
    View publication on PubMed
Search for more literature for 1.4.1.4

Cloned(Commentary)

Cloned (Comment) Organism
gene gdhA, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Escherichia coli

Protein Variants

Protein Variants Comment Organism
C321A site-directed mutagenesis, the mutant shows reduced NADP-related GDH activity, the mutation causes a greater than a 2fold drop in Vmax and more than a 2fold increase in Km for NADP+ Escherichia coli
K341L site-directed mutagenesis, the mutant shows reduced NADP-related GDH activity compared to wild-type Escherichia coli
P320A site-directed mutagenesis, the mutant shows reduced NADP-related GDH activity, the mutation causes a greater than a 2fold drop in Vmax and more than a 2fold increase in Km for NADP+. The mutant is not inhibited by propylselen Escherichia coli

Inhibitors

Inhibitors Comment Organism Structure
ebselen
-
 Escherichia coli
epigallocatechingallate EGCG Escherichia coli
propylselen inhibits glutamate dehydrogenase by binding at the NADP+ binding site. No inhibition of enzyme mutant P320A Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.07
-
 NADP+ pH 8.0, 22°C, recombinant wild-type enzyme Escherichia coli
0.13
-
 NADP+ pH 8.0, 22°C, recombinant mutant K341L Escherichia coli
0.15
-
 NADP+ pH 8.0, 22°C, recombinant mutant C321A Escherichia coli
0.24
-
 L-glutamate pH 8.0, 22°C, recombinant mutant K341L Escherichia coli
0.3
-
 NADP+ pH 8.0, 22°C, recombinant mutant P320A Escherichia coli
0.45
-
 L-glutamate pH 8.0, 22°C, recombinant wild-type enzyme Escherichia coli
0.6
-
 L-glutamate pH 8.0, 22°C, recombinant mutant C321A Escherichia coli
0.9
-
 L-glutamate pH 8.0, 22°C, recombinant mutant P320A Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-glutamate + H2O + NADP+ Escherichia coli
-
 2-oxoglutarate + NH3 + NADPH + H+
-
 r

Organism

Organism UniProt Comment Textmining
Escherichia coli P00370
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-glutamate + H2O + NADP+
-
 Escherichia coli 2-oxoglutarate + NH3 + NADPH + H+
-
 r

Subunits

Subunits Comment Organism
homohexamer
-
 Escherichia coli

Synonyms

Synonyms Comment Organism
GDH
-
 Escherichia coli
GdhA
-
 Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
22
-
 assay at room temperature Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
6.7
-
 L-glutamate pH 8.0, 22°C, recombinant mutant C321A Escherichia coli
8.8
-
 NADP+ pH 8.0, 22°C, recombinant mutant C321A Escherichia coli
9.2
-
 NADP+ pH 8.0, 22°C, recombinant mutant P320A Escherichia coli
9.6
-
 L-glutamate pH 8.0, 22°C, recombinant mutant P320A Escherichia coli
16.7
-
 NADP+ pH 8.0, 22°C, recombinant wild-type enzyme Escherichia coli
17.1
-
 L-glutamate pH 8.0, 22°C, recombinant wild-type enzyme Escherichia coli
19.6
-
 L-glutamate pH 8.0, 22°C, recombinant mutant K341L Escherichia coli
20.4
-
 NADP+ pH 8.0, 22°C, recombinant mutant K341L Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
 assay at Escherichia coli

Cofactor

Cofactor Comment Organism Structure
NADP+ P320 and C321 are both important for NADP+ binding Escherichia coli
NADPH
-
 Escherichia coli

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
additional information
-
 additional information inhibition kinetics for propylselen, ebselen, and EGCG, overview Escherichia coli

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
0.000202
-
 pH 8.0, 22°C, recombinant mutant C321A, 2.5 mm Glu Escherichia coli epigallocatechingallate
0.000202
-
 pH 8.0, 22°C, recombinant mutant P320A, 2.5 mm Glu Escherichia coli epigallocatechingallate
0.000204
-
 pH 8.0, 22°C, recombinant wild-type enzyme, at 2.5 mm Glu Escherichia coli epigallocatechingallate
0.00026
-
 pH 8.0, 22°C, recombinant mutant K341L, 2.5 mm Glu Escherichia coli ebselen
0.000333
-
 pH 8.0, 22°C, recombinant mutant K341L, 2.5 mm Glu Escherichia coli epigallocatechingallate
0.000469
-
 pH 8.0, 22°C, recombinant wild-type enzyme, 2.5 mm Glu Escherichia coli propylselen
0.0005
-
 pH 8.0, 22°C, recombinant wild-type enzyme, 2.5 mm Glu Escherichia coli ebselen
0.000585
-
 pH 8.0, 22°C, recombinant mutant K341L, 2.5 mm Glu Escherichia coli propylselen
0.0016
-
 pH 8.0, 22°C, recombinant mutant C321A, 2.5 mm Glu Escherichia coli ebselen
0.00189
-
 pH 8.0, 22°C, recombinant mutant C321A, 2.5 mm Glu Escherichia coli propylselen
0.00274
-
 pH 8.0, 22°C, recombinant mutant P320A, 2.5 mm Glu Escherichia coli ebselen

General Information

General Information Comment Organism
malfunction propylselen inhibits cancer cell growth by targeting glutamate dehydrogenase at the NADP+ binding site Escherichia coli
additional information P320 and C321 are both important for NADP+ binding Escherichia coli

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
10.67
-
 L-glutamate pH 8.0, 22°C, recombinant mutant P320A Escherichia coli
11.17
-
 L-glutamate pH 8.0, 22°C, recombinant mutant C321A Escherichia coli
30.67
-
 NADP+ pH 8.0, 22°C, recombinant mutant P320A Escherichia coli
38
-
 L-glutamate pH 8.0, 22°C, recombinant wild-type enzyme Escherichia coli
58.67
-
 NADP+ pH 8.0, 22°C, recombinant mutant C321A Escherichia coli
81.67
-
 L-glutamate pH 8.0, 22°C, recombinant mutant K341L Escherichia coli
156.9
-
 NADP+ pH 8.0, 22°C, recombinant mutant K341L Escherichia coli
238.6
-
 NADP+ pH 8.0, 22°C, recombinant wild-type enzyme Escherichia coli