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Literature summary for 1.4.1.2 extracted from

  • Engel, P.
    Glutamate dehydrogenases: the why and how of coenzyme specificity (2013), Neurochem. Res., 39, 426-432.
    View publication on PubMed

Protein Variants

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 [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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Peptoniphilus asaccharolyticus
-
-
-
[Clostridium] symbiosum
-
-
-

Substrates and Products (Substrate)

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

Subunits Comment Organism
hexamer 4 * 48000, about, SDS-PAGE Peptoniphilus asaccharolyticus
hexamer 4 * 48000, about, SDS-PAGE [Clostridium] symbiosum

Synonyms

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

Cofactor Comment Organism Structure
NAD+
-
Peptoniphilus asaccharolyticus
NAD+
-
[Clostridium] symbiosum
NADH
-
Peptoniphilus asaccharolyticus
NADH
-
[Clostridium] symbiosum

General Information

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 [mM/s]

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