BRENDA - Enzyme Database

Thr373, Asp375, and Lys260 are in the coenzyme site of porcine NADP-dependent isocitrate dehydrogenase

Lee, P.; Colman, R.F.; Arch. Biochem. Biophys. 450, 183-190 (2006)

Data extracted from this reference:

Engineering
EC Number
Amino acid exchange
Commentary
Organism
1.1.1.42
D375N
15fold increase in KM-value for NADP+, marked decrease of Vmax-value
Sus scrofa
1.1.1.42
K260Q
28fold increase in KM-value for NADP+, marked decrease of Vmax-value
Sus scrofa
1.1.1.42
K374Q
little change in kinetic parameters
Sus scrofa
1.1.1.42
T373A
reduction of Vmax-value to 1% of wild-type
Sus scrofa
1.1.1.42
T373S
little change in kinetic parameters
Sus scrofa
1.1.1.42
T373V
reduction of Vmax-value to 20% of wild-type
Sus scrofa
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.42
0.0055
-
NADP+
mutant T373S, 25°C, pH 7.4
Sus scrofa
1.1.1.42
0.0062
-
NADP+
mutant T373A, 25°C, pH 7.4
Sus scrofa
1.1.1.42
0.0088
-
NADP+
wild-type, 25°C, pH 7.4
Sus scrofa
1.1.1.42
0.012
-
NADP+
mutant T373V, 25°C, pH 7.4
Sus scrofa
1.1.1.42
0.0154
-
NADP+
mutant K374Q 25°C, pH 7.4
Sus scrofa
1.1.1.42
0.0179
-
NAD+
wild-type, 25°C, pH 7.4
Sus scrofa
1.1.1.42
0.0207
-
NAD+
mutant K260R, 25°C, pH 7.4
Sus scrofa
1.1.1.42
0.0232
-
NAD+
mutant D375N, 25°C, pH 7.4
Sus scrofa
1.1.1.42
0.0312
-
NAD+
mutant K260Q, 25°C, pH 7.4
Sus scrofa
1.1.1.42
0.0378
-
NADP+
mutant K260R, 25°C, pH 7.4
Sus scrofa
1.1.1.42
0.133
-
NADP+
mutant D375N, 25°C, pH 7.4
Sus scrofa
1.1.1.42
0.25
-
NADP+
mutant K260Q, 25°C, pH 7.4
Sus scrofa
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.1.1.42
Sus scrofa
P33198
-
-
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.1.1.42
DL-isocitrate + NAD+
-
667326
Sus scrofa
2-oxoglutarate + CO2 + NADH
-
-
-
?
1.1.1.42
DL-isocitrate + NADP+
-
667326
Sus scrofa
2-oxoglutarate + CO2 + NADPH
-
-
-
?
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.1.1.42
0.03
-
NAD+
mutant D375N, 25°C, pH 7.4
Sus scrofa
1.1.1.42
0.47
-
NAD+
mutant K260Q, 25°C, pH 7.4
Sus scrofa
1.1.1.42
0.5
-
NAD+
mutant K260R, 25°C, pH 7.4
Sus scrofa
1.1.1.42
0.76
-
NAD+
wild-type, 25°C, pH 7.4
Sus scrofa
1.1.1.42
1.7
-
NADP+
mutant K260Q, 25°C, pH 7.4
Sus scrofa
1.1.1.42
5.7
-
NADP+
mutant K260R, 25°C, pH 7.4
Sus scrofa
1.1.1.42
8.4
-
NADP+
mutant D375N, 25°C, pH 7.4
Sus scrofa
1.1.1.42
32.2
-
NADP+
wild-type, 25°C, pH 7.4
Sus scrofa
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
1.1.1.42
D375N
15fold increase in KM-value for NADP+, marked decrease of Vmax-value
Sus scrofa
1.1.1.42
K260Q
28fold increase in KM-value for NADP+, marked decrease of Vmax-value
Sus scrofa
1.1.1.42
K374Q
little change in kinetic parameters
Sus scrofa
1.1.1.42
T373A
reduction of Vmax-value to 1% of wild-type
Sus scrofa
1.1.1.42
T373S
little change in kinetic parameters
Sus scrofa
1.1.1.42
T373V
reduction of Vmax-value to 20% of wild-type
Sus scrofa
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.42
0.0055
-
NADP+
mutant T373S, 25°C, pH 7.4
Sus scrofa
1.1.1.42
0.0062
-
NADP+
mutant T373A, 25°C, pH 7.4
Sus scrofa
1.1.1.42
0.0088
-
NADP+
wild-type, 25°C, pH 7.4
Sus scrofa
1.1.1.42
0.012
-
NADP+
mutant T373V, 25°C, pH 7.4
Sus scrofa
1.1.1.42
0.0154
-
NADP+
mutant K374Q 25°C, pH 7.4
Sus scrofa
1.1.1.42
0.0179
-
NAD+
wild-type, 25°C, pH 7.4
Sus scrofa
1.1.1.42
0.0207
-
NAD+
mutant K260R, 25°C, pH 7.4
Sus scrofa
1.1.1.42
0.0232
-
NAD+
mutant D375N, 25°C, pH 7.4
Sus scrofa
1.1.1.42
0.0312
-
NAD+
mutant K260Q, 25°C, pH 7.4
Sus scrofa
1.1.1.42
0.0378
-
NADP+
mutant K260R, 25°C, pH 7.4
Sus scrofa
1.1.1.42
0.133
-
NADP+
mutant D375N, 25°C, pH 7.4
Sus scrofa
1.1.1.42
0.25
-
NADP+
mutant K260Q, 25°C, pH 7.4
Sus scrofa
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.1.1.42
DL-isocitrate + NAD+
-
667326
Sus scrofa
2-oxoglutarate + CO2 + NADH
-
-
-
?
1.1.1.42
DL-isocitrate + NADP+
-
667326
Sus scrofa
2-oxoglutarate + CO2 + NADPH
-
-
-
?
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.1.1.42
0.03
-
NAD+
mutant D375N, 25°C, pH 7.4
Sus scrofa
1.1.1.42
0.47
-
NAD+
mutant K260Q, 25°C, pH 7.4
Sus scrofa
1.1.1.42
0.5
-
NAD+
mutant K260R, 25°C, pH 7.4
Sus scrofa
1.1.1.42
0.76
-
NAD+
wild-type, 25°C, pH 7.4
Sus scrofa
1.1.1.42
1.7
-
NADP+
mutant K260Q, 25°C, pH 7.4
Sus scrofa
1.1.1.42
5.7
-
NADP+
mutant K260R, 25°C, pH 7.4
Sus scrofa
1.1.1.42
8.4
-
NADP+
mutant D375N, 25°C, pH 7.4
Sus scrofa
1.1.1.42
32.2
-
NADP+
wild-type, 25°C, pH 7.4
Sus scrofa