BRENDA - Enzyme Database

The role of glutamate 87 in the kinetic mechanism of Thermus thermophilus isopropylmalate dehydrogenase

Dean, A.M.; Dvorak, L.; Protein Sci. 4, 2156-2167 (1995)

Data extracted from this reference:

Engineering
EC Number
Amino acid exchange
Commentary
Organism
1.1.1.85
E87G
mutation does not affect the dissociation constant from the binary complex, but does greatly increase the KM-values
Thermus thermophilus
1.1.1.85
E87Q
mutation does not affect the dissociation constant from the binary complex, but does greatly increase the KM-values
Thermus thermophilus
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
1.1.1.85
isocitrate
-
Thermus thermophilus
1.1.1.85
NADH
NADH always produces competitive inhibition patterns with respect to NAD and noncompetitive inhibition patterns with respect to the substrates
Thermus thermophilus
1.1.1.85
Tartrate
-
Thermus thermophilus
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.85
0.000017
-
isopropylmalate
pH 7.3, 21°C, cofactor: NAD+, wild-type enzyme
Thermus thermophilus
1.1.1.85
0.00048
-
isopropylmalate
pH 7.3, 21°C, cofactor: NAD+, mutant enzyme E87G
Thermus thermophilus
1.1.1.85
0.00298
-
isopropylmalate
pH 7.3, 21°C, cofactor: NADP+, wild-type enzyme
Thermus thermophilus
1.1.1.85
0.00321
-
NAD+
pH 7.3, 21°C, reaction with isopropylmalate, wild-type enzyme
Thermus thermophilus
1.1.1.85
0.0053
-
isopropylmalate
pH 7.3, 21°C, cofactor: NAD+, mutant enzyme E87Q
Thermus thermophilus
1.1.1.85
0.0326
-
malate
pH 7.3, 21°C, cofactor: NAD+, isopropylmalate dehydrogenase mutant E87Q
Thermus thermophilus
1.1.1.85
0.0407
-
Tartrate
pH 7.3, 21°C, cofactor: NAD+, wild-type enzyme
Thermus thermophilus
1.1.1.85
0.0454
-
NAD+
pH 7.3, 21°C, reaction with tartrate, wild-type enzym
Thermus thermophilus
1.1.1.85
0.1065
-
NAD+
pH 7.3, 21°C, reaction with malate, wild-type enzyme
Thermus thermophilus
1.1.1.85
0.131
-
NAD+
pH 7.3, 21°C, isopropylmalate dehydrogenase mutant E87G
Thermus thermophilus
1.1.1.85
0.181
-
NAD+
pH 7.3, 21°C, isopropylmalate dehydrogenase mutant E87Q
Thermus thermophilus
1.1.1.85
0.386
-
NAD+
pH 7.3, 21°C, isopropylmalate dehydrogenase mutant E87G
Thermus thermophilus
1.1.1.85
0.761
-
Tartrate
pH 7.3, 21°C, cofactor: NAD+, isopropylmalate dehydrogenase mutant E87Q
Thermus thermophilus
1.1.1.85
0.778
-
NAD+
pH 7.3, 21°C, isopropylmalate dehydrogenase mutant E87Q
Thermus thermophilus
1.1.1.85
0.817
-
Tartrate
pH 7.3, 21°C, cofactor: NADP+, wild-type enzyme
Thermus thermophilus
1.1.1.85
1.21
-
Tartrate
pH 7.3, 21°C, cofactor: NAD+, mutant enzyme E87G
Thermus thermophilus
1.1.1.85
1.51
-
malate
pH 7.3, 21°C, cofactor: NAD+, wild-type enzyme
Thermus thermophilus
1.1.1.85
1.573
-
NADP+
pH 7.3, 21°C, reaction with tartrate, wild-type enzyme
Thermus thermophilus
1.1.1.85
1.836
-
NADP+
pH 7.3, 21°C, reaction with isopropylmalate, wild-type enzyme
Thermus thermophilus
1.1.1.85
1.853
-
NAD+
pH 7.3, 21°C, reaction with malate, mutant enzyme E87Q
Thermus thermophilus
1.1.1.85
2.584
-
NAD+
pH 7.3, 21°C, reaction with malate, mutant enzyme E87G
Thermus thermophilus
1.1.1.85
6.579
-
NADP+
pH 7.3, 21°C, recation with malate, wild-type enzyme
Thermus thermophilus
1.1.1.85
31.56
-
malate
pH 7.3, 21°C, cofactor: NADP+, wild-type enzyme
Thermus thermophilus
1.1.1.85
35.59
-
malate
pH 7.3, 21°C, cofactor: NAD+, isopropylmalate dehydrogenase mutant E87G
Thermus thermophilus
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.1.1.85
Thermus thermophilus
-
-
-
Reaction
EC Number
Reaction
Commentary
Organism
1.1.1.85
(2R,3S)-3-isopropylmalate + NAD+ = (2S)-2-isopropyl-3-oxosuccinate + NADH + H+
the oxidative decarboxylation of 2-hydroxy acid substrates by the enzyme occurs via a random kinetic mechanism with the formation of two abortive complexes: enzyme-NADH-substrate and enzyme-NADH-inhibitor
Thermus thermophilus
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.1.1.85
(2R,3S)-3-isopropylmalate + NAD+
-
639155
Thermus thermophilus
2-oxoisocaproate + NADH + CO2
-
-
-
?
1.1.1.85
malate + NAD+
-
639155
Thermus thermophilus
pyruvate + NADH + CO2
-
-
-
?
1.1.1.85
malate + NADP+
-
639155
Thermus thermophilus
pyruvate + NADPH + CO2
-
-
-
?
1.1.1.85
tartrate + NAD+
-
639155
Thermus thermophilus
?
-
-
-
?
1.1.1.85
tartrate + NADP+
-
639155
Thermus thermophilus
?
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
1.1.1.85
dimer
2 * 35000-40000, SDS-PAGE
Thermus thermophilus
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.1.1.85
0.1
-
NADP+
pH 7.3, 21°C, reaction with tartrate, wild-type enzyme
Thermus thermophilus
1.1.1.85
0.1
-
Tartrate
pH 7.3, 21°C, reaction with NADP+, wild-type enzyme
Thermus thermophilus
1.1.1.85
0.19
-
malate
pH 7.3, 21°C, reaction with NAD+, mutant enzyme E87Q
Thermus thermophilus
1.1.1.85
0.19
-
NAD+
pH 7.3, 21°C, reaction with tartrate, wild-type enzyme or reaction with malate, mutant enzyme E87Q
Thermus thermophilus
1.1.1.85
0.19
-
Tartrate
pH 7.3, 21°C, reaction with NAD+, wild-type enzyme
Thermus thermophilus
1.1.1.85
0.2
-
isopropylmalate
pH 7.3, 21°C, reaction with NAD+, mutant enzyme E87G
Thermus thermophilus
1.1.1.85
0.2
-
NAD+
pH 7.3, 21°C, reaction with isopropylmalate, mutant enzyme E87G
Thermus thermophilus
1.1.1.85
0.21
-
isopropylmalate
pH 7.3, 21°C, reaction with NAD+, mutant enzyme E87Q
Thermus thermophilus
1.1.1.85
0.21
-
NAD+
pH 7.3, 21°C, reaction with isopropylmalate, mutant enzyme E87Q; pH 7.3, 21°C, reaction with tartrate, mutant enzyme E87G
Thermus thermophilus
1.1.1.85
0.21
-
Tartrate
pH 7.3, 21°C, reaction with NAD+, mutant enzyme E87G
Thermus thermophilus
1.1.1.85
0.24
-
isopropylmalate
pH 7.3, 21°C, reaction with NAD+, wild-type enzyme
Thermus thermophilus
1.1.1.85
0.24
-
NAD+
pH 7.3, 21°C, reaction with isopropylmalate, wild-type enzyme
Thermus thermophilus
1.1.1.85
0.26
-
malate
pH 7.3, 21°C, reaction with NAD+, mutant enzyme E87G
Thermus thermophilus
1.1.1.85
0.26
-
NAD+
pH 7.3, 21°C, reaction with malate, mutant enzyme E87G or reaction with tartrate, mutant enzyme E87Q
Thermus thermophilus
1.1.1.85
0.26
-
Tartrate
pH 7.3, 21°C, reaction with NAD+, mutant enzyme E87Q
Thermus thermophilus
1.1.1.85
1.66
-
isopropylmalate
pH 7.3, 21°C, reaction with NADP+, wild-type enzyme
Thermus thermophilus
1.1.1.85
1.66
-
NADP+
pH 7.3, 21°C, reaction with isopropylmalate, wild-type enzyme
Thermus thermophilus
1.1.1.85
4.69
-
malate
pH 7.3, 21°C, reaction with NADP+, wild-type enzyme
Thermus thermophilus
1.1.1.85
4.69
-
NADP+
pH 7.3, 21°C, reaction with malate, wild-type enzyme
Thermus thermophilus
1.1.1.85
10.6
-
malate
pH 7.3, 21°C, reaction with NAD+, wild-type enzyme
Thermus thermophilus
1.1.1.85
10.6
-
NAD+
pH 7.3, 21°C, reaction with malate, wild-type enzyme
Thermus thermophilus
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.85
NAD+
-
Thermus thermophilus
1.1.1.85
NADP+
-
Thermus thermophilus
Ki Value [mM]
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
1.1.1.85
additional information
-
additional information
-
Thermus thermophilus
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.85
NAD+
-
Thermus thermophilus
1.1.1.85
NADP+
-
Thermus thermophilus
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
1.1.1.85
E87G
mutation does not affect the dissociation constant from the binary complex, but does greatly increase the KM-values
Thermus thermophilus
1.1.1.85
E87Q
mutation does not affect the dissociation constant from the binary complex, but does greatly increase the KM-values
Thermus thermophilus
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
1.1.1.85
isocitrate
-
Thermus thermophilus
1.1.1.85
NADH
NADH always produces competitive inhibition patterns with respect to NAD and noncompetitive inhibition patterns with respect to the substrates
Thermus thermophilus
1.1.1.85
Tartrate
-
Thermus thermophilus
Ki Value [mM] (protein specific)
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
1.1.1.85
additional information
-
additional information
-
Thermus thermophilus
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.85
0.000017
-
isopropylmalate
pH 7.3, 21°C, cofactor: NAD+, wild-type enzyme
Thermus thermophilus
1.1.1.85
0.00048
-
isopropylmalate
pH 7.3, 21°C, cofactor: NAD+, mutant enzyme E87G
Thermus thermophilus
1.1.1.85
0.00298
-
isopropylmalate
pH 7.3, 21°C, cofactor: NADP+, wild-type enzyme
Thermus thermophilus
1.1.1.85
0.00321
-
NAD+
pH 7.3, 21°C, reaction with isopropylmalate, wild-type enzyme
Thermus thermophilus
1.1.1.85
0.0053
-
isopropylmalate
pH 7.3, 21°C, cofactor: NAD+, mutant enzyme E87Q
Thermus thermophilus
1.1.1.85
0.0326
-
malate
pH 7.3, 21°C, cofactor: NAD+, isopropylmalate dehydrogenase mutant E87Q
Thermus thermophilus
1.1.1.85
0.0407
-
Tartrate
pH 7.3, 21°C, cofactor: NAD+, wild-type enzyme
Thermus thermophilus
1.1.1.85
0.0454
-
NAD+
pH 7.3, 21°C, reaction with tartrate, wild-type enzym
Thermus thermophilus
1.1.1.85
0.1065
-
NAD+
pH 7.3, 21°C, reaction with malate, wild-type enzyme
Thermus thermophilus
1.1.1.85
0.131
-
NAD+
pH 7.3, 21°C, isopropylmalate dehydrogenase mutant E87G
Thermus thermophilus
1.1.1.85
0.181
-
NAD+
pH 7.3, 21°C, isopropylmalate dehydrogenase mutant E87Q
Thermus thermophilus
1.1.1.85
0.386
-
NAD+
pH 7.3, 21°C, isopropylmalate dehydrogenase mutant E87G
Thermus thermophilus
1.1.1.85
0.761
-
Tartrate
pH 7.3, 21°C, cofactor: NAD+, isopropylmalate dehydrogenase mutant E87Q
Thermus thermophilus
1.1.1.85
0.778
-
NAD+
pH 7.3, 21°C, isopropylmalate dehydrogenase mutant E87Q
Thermus thermophilus
1.1.1.85
0.817
-
Tartrate
pH 7.3, 21°C, cofactor: NADP+, wild-type enzyme
Thermus thermophilus
1.1.1.85
1.21
-
Tartrate
pH 7.3, 21°C, cofactor: NAD+, mutant enzyme E87G
Thermus thermophilus
1.1.1.85
1.51
-
malate
pH 7.3, 21°C, cofactor: NAD+, wild-type enzyme
Thermus thermophilus
1.1.1.85
1.573
-
NADP+
pH 7.3, 21°C, reaction with tartrate, wild-type enzyme
Thermus thermophilus
1.1.1.85
1.836
-
NADP+
pH 7.3, 21°C, reaction with isopropylmalate, wild-type enzyme
Thermus thermophilus
1.1.1.85
1.853
-
NAD+
pH 7.3, 21°C, reaction with malate, mutant enzyme E87Q
Thermus thermophilus
1.1.1.85
2.584
-
NAD+
pH 7.3, 21°C, reaction with malate, mutant enzyme E87G
Thermus thermophilus
1.1.1.85
6.579
-
NADP+
pH 7.3, 21°C, recation with malate, wild-type enzyme
Thermus thermophilus
1.1.1.85
31.56
-
malate
pH 7.3, 21°C, cofactor: NADP+, wild-type enzyme
Thermus thermophilus
1.1.1.85
35.59
-
malate
pH 7.3, 21°C, cofactor: NAD+, isopropylmalate dehydrogenase mutant E87G
Thermus thermophilus
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.1.1.85
(2R,3S)-3-isopropylmalate + NAD+
-
639155
Thermus thermophilus
2-oxoisocaproate + NADH + CO2
-
-
-
?
1.1.1.85
malate + NAD+
-
639155
Thermus thermophilus
pyruvate + NADH + CO2
-
-
-
?
1.1.1.85
malate + NADP+
-
639155
Thermus thermophilus
pyruvate + NADPH + CO2
-
-
-
?
1.1.1.85
tartrate + NAD+
-
639155
Thermus thermophilus
?
-
-
-
?
1.1.1.85
tartrate + NADP+
-
639155
Thermus thermophilus
?
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.1.1.85
dimer
2 * 35000-40000, SDS-PAGE
Thermus thermophilus
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.1.1.85
0.1
-
NADP+
pH 7.3, 21°C, reaction with tartrate, wild-type enzyme
Thermus thermophilus
1.1.1.85
0.1
-
Tartrate
pH 7.3, 21°C, reaction with NADP+, wild-type enzyme
Thermus thermophilus
1.1.1.85
0.19
-
malate
pH 7.3, 21°C, reaction with NAD+, mutant enzyme E87Q
Thermus thermophilus
1.1.1.85
0.19
-
NAD+
pH 7.3, 21°C, reaction with tartrate, wild-type enzyme or reaction with malate, mutant enzyme E87Q
Thermus thermophilus
1.1.1.85
0.19
-
Tartrate
pH 7.3, 21°C, reaction with NAD+, wild-type enzyme
Thermus thermophilus
1.1.1.85
0.2
-
isopropylmalate
pH 7.3, 21°C, reaction with NAD+, mutant enzyme E87G
Thermus thermophilus
1.1.1.85
0.2
-
NAD+
pH 7.3, 21°C, reaction with isopropylmalate, mutant enzyme E87G
Thermus thermophilus
1.1.1.85
0.21
-
isopropylmalate
pH 7.3, 21°C, reaction with NAD+, mutant enzyme E87Q
Thermus thermophilus
1.1.1.85
0.21
-
NAD+
pH 7.3, 21°C, reaction with isopropylmalate, mutant enzyme E87Q; pH 7.3, 21°C, reaction with tartrate, mutant enzyme E87G
Thermus thermophilus
1.1.1.85
0.21
-
Tartrate
pH 7.3, 21°C, reaction with NAD+, mutant enzyme E87G
Thermus thermophilus
1.1.1.85
0.24
-
isopropylmalate
pH 7.3, 21°C, reaction with NAD+, wild-type enzyme
Thermus thermophilus
1.1.1.85
0.24
-
NAD+
pH 7.3, 21°C, reaction with isopropylmalate, wild-type enzyme
Thermus thermophilus
1.1.1.85
0.26
-
malate
pH 7.3, 21°C, reaction with NAD+, mutant enzyme E87G
Thermus thermophilus
1.1.1.85
0.26
-
NAD+
pH 7.3, 21°C, reaction with malate, mutant enzyme E87G or reaction with tartrate, mutant enzyme E87Q
Thermus thermophilus
1.1.1.85
0.26
-
Tartrate
pH 7.3, 21°C, reaction with NAD+, mutant enzyme E87Q
Thermus thermophilus
1.1.1.85
1.66
-
isopropylmalate
pH 7.3, 21°C, reaction with NADP+, wild-type enzyme
Thermus thermophilus
1.1.1.85
1.66
-
NADP+
pH 7.3, 21°C, reaction with isopropylmalate, wild-type enzyme
Thermus thermophilus
1.1.1.85
4.69
-
malate
pH 7.3, 21°C, reaction with NADP+, wild-type enzyme
Thermus thermophilus
1.1.1.85
4.69
-
NADP+
pH 7.3, 21°C, reaction with malate, wild-type enzyme
Thermus thermophilus
1.1.1.85
10.6
-
malate
pH 7.3, 21°C, reaction with NAD+, wild-type enzyme
Thermus thermophilus
1.1.1.85
10.6
-
NAD+
pH 7.3, 21°C, reaction with malate, wild-type enzyme
Thermus thermophilus