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Literature summary for 1.4.1.4 extracted from

  • Tomita, T.; Yin, L.; Nakamura, S.; Kosono, S.; Kuzuyama, T.; Nishiyama, M.
    Crystal structure of the 2-iminoglutarate-bound complex of glutamate dehydrogenase from Corynebacterium glutamicum (2017), FEBS Lett., 591, 1611-1622 .
    View publication on PubMed
Search for more literature for 1.4.1.4

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Corynebacterium glutamicum

Crystallization (Commentary)

Crystallization (Comment) Organism
complexed with NADP+ and 2-iminoglutarate. Among six subunits of hexameric GDH-binding NADP+, only four subunits bind 2-iminoglutarate in a closed form, while the other two are in an open form. In the closed form, 2-iminoglutarate is bound to the substrate-binding site with the 2-imino group stacked by the nicotinamide ring of the coenzyme, suggesting a prehydride transfer state Corynebacterium glutamicum

Protein Variants

Protein Variants Comment Organism
K136A residue is directly involved in binding the 2'-phosphate group of NADP+, increase in Km value for NADPH, fourfold increase in the Km value for NADH with a concomitant 1.6fold increase in the kcat value Corynebacterium glutamicum
R290A residue is directly involved in binding the 2'-phosphate group of NADP+, increase in Km value for NADPH Corynebacterium glutamicum
S265A residue is directly involved in binding the 2'-phosphate group of NADP+, increase in Km value for NADPH Corynebacterium glutamicum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.016
-
 NADPH wild-type, pH 7.0,30°C Corynebacterium glutamicum
0.058
-
 NADPH mutant K1376A, pH 7.0,30°C Corynebacterium glutamicum
0.073
-
 NADPH mutant R290A, pH 7.0,30°C Corynebacterium glutamicum
0.093
-
 NADP+ wild-type, pH 7.0,30°C Corynebacterium glutamicum
0.16
-
 NADPH mutant S265A, pH 7.0,30°C Corynebacterium glutamicum
0.16
-
 NADH wild-type, pH 7.0,30°C Corynebacterium glutamicum
0.69
-
 NADH mutant K136A, pH 7.0,30°C Corynebacterium glutamicum
0.89
-
 2-oxoglutarate wild-type, pH 7.0,30°C Corynebacterium glutamicum
33
-
 NH3 wild-type, pH 7.0,30°C Corynebacterium glutamicum
50
-
 L-glutamate wild-type, pH 7.0,30°C Corynebacterium glutamicum

Organism

Organism UniProt Comment Textmining
Corynebacterium glutamicum A0A0U4QBJ6
-
-
Corynebacterium glutamicum ATCC 13869 A0A0U4QBJ6
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-oxoglutarate + NH3 + NADH + H+
-
 Corynebacterium glutamicum L-glutamate + H2O + NAD+
-
 r
2-oxoglutarate + NH3 + NADH + H+
-
 Corynebacterium glutamicum ATCC 13869 L-glutamate + H2O + NAD+
-
 r
2-oxoglutarate + NH3 + NADPH + H+
-
 Corynebacterium glutamicum L-glutamate + H2O + NADP+
-
 r
2-oxoglutarate + NH3 + NADPH + H+
-
 Corynebacterium glutamicum ATCC 13869 L-glutamate + H2O + NADP+
-
 r
L-glutamate + H2O + NADP+
-
 Corynebacterium glutamicum 2-oxoglutarate + NH3 + NADPH + H+
-
 r
L-glutamate + H2O + NADP+
-
 Corynebacterium glutamicum ATCC 13869 2-oxoglutarate + NH3 + NADPH + H+
-
 r

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.4
-
 NADH wild-type, pH 7.0,30°C Corynebacterium glutamicum
2.3
-
 NADH mutant K136A, pH 7.0,30°C Corynebacterium glutamicum
8.1
-
 NADP+ wild-type, pH 7.0,30°C Corynebacterium glutamicum
9.3
-
 L-glutamate wild-type, pH 7.0,30°C Corynebacterium glutamicum
1900
-
 NADPH mutant K1376A, pH 7.0,30°C Corynebacterium glutamicum
2200
-
 NADPH wild-type, pH 7.0,30°C Corynebacterium glutamicum
2500
-
 2-oxoglutarate wild-type, pH 7.0,30°C Corynebacterium glutamicum
3000
-
 NADPH mutant R290A, pH 7.0,30°C Corynebacterium glutamicum
3300
-
 NADPH mutant S265A, pH 7.0,30°C Corynebacterium glutamicum
3400
-
 NH3 wild-type, pH 7.0,30°C Corynebacterium glutamicum

Cofactor

Cofactor Comment Organism Structure
NADH NADPH is 10000fold preferred over NADH Corynebacterium glutamicum
NADPH
-
 Corynebacterium glutamicum

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.19
-
 L-glutamate wild-type, pH 7.0,30°C Corynebacterium glutamicum
3.3
-
 NADH mutant K136A, pH 7.0,30°C Corynebacterium glutamicum
8.8
-
 NADH wild-type, pH 7.0,30°C Corynebacterium glutamicum
87
-
 NADP+ wild-type, pH 7.0,30°C Corynebacterium glutamicum
100
-
 NH3 wild-type, pH 7.0,30°C Corynebacterium glutamicum
2800
-
 2-oxoglutarate wild-type, pH 7.0,30°C Corynebacterium glutamicum
21000
-
 NADPH mutant S265A, pH 7.0,30°C Corynebacterium glutamicum
34000
-
 NADPH mutant K1376A, pH 7.0,30°C Corynebacterium glutamicum
42000
-
 NADPH mutant R290A, pH 7.0,30°C Corynebacterium glutamicum
130000
-
 NADPH wild-type, pH 7.0,30°C Corynebacterium glutamicum