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 | 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 | 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 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 | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-glutamate + H2O + NADP+ | Escherichia coli | - |
2-oxoglutarate + NH3 + NADPH + H+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P00370 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-glutamate + H2O + NADP+ | - |
Escherichia coli | 2-oxoglutarate + NH3 + NADPH + H+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
homohexamer | - |
Escherichia coli |
Synonyms | Comment | Organism |
---|---|---|
GDH | - |
Escherichia coli |
GdhA | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
22 | - |
assay at room temperature | Escherichia coli |
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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADP+ | P320 and C321 are both important for NADP+ binding | Escherichia coli | |
NADPH | - |
Escherichia coli |
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 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 | 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 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 |