Protein Variants | Comment | Organism |
---|---|---|
D245K | site-directed mutagenesis, discrimination against NADPH by factor 32, compared to 1000 for the wild-type enzyme | Peptoniphilus asaccharolyticus |
E243D | site-directed mutagenesis, substitution of Asp for Glu in E243D produces a shift in favour of NADPH by virtue of a threefold increase in the Km for NAD+ and a threefold decrease in that for NADP+, resulting in a 9fold shift in the overall discrimination factor, discrimination against NADPH by factor 130, compared to 1000 for the wild-type enzyme | Peptoniphilus asaccharolyticus |
E243K | site-directed mutagenesis, discrimination against NADPH by a factor below 130, compared to 1000 for the wild-type enzyme | Peptoniphilus asaccharolyticus |
F232S/P262S/D263K | site-directed mutagenesis, the mutant shows switched cofactor spcificity compared to the wild-type enzyme, it has high activity with NADPH/NADP+ | [Clostridium] symbiosum |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
48000 | - |
4 * 48000, about, SDS-PAGE | Peptoniphilus asaccharolyticus |
48000 | - |
4 * 48000, about, SDS-PAGE | [Clostridium] symbiosum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-glutamate + H2O + NAD+ | Peptoniphilus asaccharolyticus | - |
2-oxoglutarate + NH3 + NADH + H+ | - |
r | |
L-glutamate + H2O + NAD+ | [Clostridium] symbiosum | - |
2-oxoglutarate + NH3 + NADH + H+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Peptoniphilus asaccharolyticus | - |
- |
- |
[Clostridium] symbiosum | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-glutamate + H2O + NAD+ | - |
Peptoniphilus asaccharolyticus | 2-oxoglutarate + NH3 + NADH + H+ | - |
r | |
L-glutamate + H2O + NAD+ | - |
[Clostridium] symbiosum | 2-oxoglutarate + NH3 + NADH + H+ | - |
r | |
L-glutamate + H2O + NAD+ | the amination reaction is preferred | [Clostridium] symbiosum | 2-oxoglutarate + NH3 + NADH + H+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
hexamer | 4 * 48000, about, SDS-PAGE | Peptoniphilus asaccharolyticus |
hexamer | 4 * 48000, about, SDS-PAGE | [Clostridium] symbiosum |
Synonyms | Comment | Organism |
---|---|---|
NAD+-dependent GDH | - |
Peptoniphilus asaccharolyticus |
NAD+-dependent GDH | - |
[Clostridium] symbiosum |
NAD+-dependent glutamate dehydrogenase | - |
Peptoniphilus asaccharolyticus |
NAD+-dependent glutamate dehydrogenase | - |
[Clostridium] symbiosum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Peptoniphilus asaccharolyticus | |
NAD+ | - |
[Clostridium] symbiosum | |
NADH | - |
Peptoniphilus asaccharolyticus | |
NADH | - |
[Clostridium] symbiosum |
General Information | Comment | Organism |
---|---|---|
evolution | NAD+-dependent, NADP+-dependent and dual-specificity GDHs, EC 1.4.1.2-1.4.1.4 are closely related and a few site-directed mutations can reverse specificity, overview. Specificity for NAD+ or for NADP+ has probably emerged repeatedly during evolution, using different structural solutions on different occasions. an acidic P7 residue usually hydrogen bonds to the 2'- and 3'-hydroxyls, may permit binding of NAD+ only, NADP+ only, or in higher animals both | Peptoniphilus asaccharolyticus |
evolution | NAD+-dependent, NADP+-dependent and dual-specificity GDHs, EC 1.4.1.2-1.4.1.4 are closely related and a few site-directed mutations can reverse specificity, overview. Specificity for NAD+ or for NADP+ has probably emerged repeatedly during evolution, using different structural solutions on different occasions. an acidic P7 residue usually hydrogen bonds to the 2'- and 3'-hydroxyls, may permit binding of NAD+ only, NADP+ only, or in higher animals both | [Clostridium] symbiosum |
additional information | GDH from Peptoniphilus asaccharolyticus obeys the rules with Gly at P6 and Glu at P7 | Peptoniphilus asaccharolyticus |
additional information | in clostridial GDH, which shows a remarkable discrimination (20000-80000fold) in favour of NAD+, the P6 residue, which should be Gly, is in fact Ala. Not only this, but the critical P7 residue is Gly instead of Asp or Glu | [Clostridium] symbiosum |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.29 | - |
NAD+ | mutant F232S/P262S/D263K , pH and temperature not specified in the publication | [Clostridium] symbiosum | |
149 | - |
NAD+ | wild-type enzyme, pH and temperature not specified in the publication | [Clostridium] symbiosum | |
369 | - |
NADH | mutant F232S/P262S/D263K , pH and temperature not specified in the publication | [Clostridium] symbiosum | |
8110 | - |
NADH | wild-type enzyme, pH and temperature not specified in the publication | [Clostridium] symbiosum |