BRENDA - Enzyme Database

Homologous binding sites in yeast isocitrate dehydrogenase for cofactor (NAD+) and allosteric activator (AMP)

Lin, A.P.; McAlister-Henn, L.; J. Biol. Chem. 278, 12864-12872 (2003)

Data extracted from this reference:

Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
1.1.1.41
AMP
allosteric activator, binding site at subunit IDH1
Saccharomyces cerevisiae
Cloned(Commentary)
EC Number
Commentary
Organism
1.1.1.41
expression of His-tagged wild-type and mutant enzymes in yeast strain IDH12DELTAL deficient in both subunits of the enzyme
Saccharomyces cerevisiae
Engineering
EC Number
Amino acid exchange
Commentary
Organism
1.1.1.41
D279A/D280A
site-directed mutagenesis, mutation of a IDH1 residue, reduced AMP binding, ligand-binding analysis
Saccharomyces cerevisiae
1.1.1.41
D286A/I287A
site-directed mutagenesis, mutation of a IDH2 residue, highly reduced NAD+ binding, ligand-binding analysis
Saccharomyces cerevisiae
1.1.1.41
H281A
site-directed mutagenesis, mutation of a IDH2 residue, mutant cannot bind NAD+, ligand-binding analysis
Saccharomyces cerevisiae
1.1.1.41
R274A
site-directed mutagenesis, mutation of a IDH1 residue, reduced AMP binding, ligand-binding analysis
Saccharomyces cerevisiae
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.41
additional information
-
additional information
kinetics and ligand binding
Saccharomyces cerevisiae
Localization
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
1.1.1.41
mitochondrion
-
Saccharomyces cerevisiae
5739
-
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
1.1.1.41
Mg2+
binding site at subunit IDH2, active site
Saccharomyces cerevisiae
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.1.1.41
303000
-
sequence calculation
Saccharomyces cerevisiae
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.1.1.41
isocitrate + NAD+
Saccharomyces cerevisiae
enzyme performs the rate-limiting step in the mitochondrial tricarboxylic cycle and is subject to complex allosteric regulation
2-oxoglutarate + CO2 + NADH
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.1.1.41
Saccharomyces cerevisiae
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
1.1.1.41
recombinant His-tagged wild-type and mutant enzymes from strain IDH12DELTAL by nickel affinity chromatography
Saccharomyces cerevisiae
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.1.1.41
D-isocitrate + NAD+
catalytic residues binding isocitrate are located on subunit IDH2, while regulatory residues binding isocitrate are located on subunit IDH1
656118
Saccharomyces cerevisiae
2-oxoglutarate + CO2 + NADH
-
-
-
?
1.1.1.41
isocitrate + NAD+
enzyme performs the rate-limiting step in the mitochondrial tricarboxylic cycle and is subject to complex allosteric regulation
656118
Saccharomyces cerevisiae
2-oxoglutarate + CO2 + NADH
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
1.1.1.41
octamer
alpha4beta4, subunits are termed IDH1 and IDH2
Saccharomyces cerevisiae
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.1.1.41
24
-
assay at
Saccharomyces cerevisiae
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.1.1.41
7.4
-
assay at
Saccharomyces cerevisiae
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.41
NAD+
specific for, binding site at subunit IDH2
Saccharomyces cerevisiae
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
1.1.1.41
AMP
allosteric activator, binding site at subunit IDH1
Saccharomyces cerevisiae
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.41
expression of His-tagged wild-type and mutant enzymes in yeast strain IDH12DELTAL deficient in both subunits of the enzyme
Saccharomyces cerevisiae
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.41
NAD+
specific for, binding site at subunit IDH2
Saccharomyces cerevisiae
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
1.1.1.41
D279A/D280A
site-directed mutagenesis, mutation of a IDH1 residue, reduced AMP binding, ligand-binding analysis
Saccharomyces cerevisiae
1.1.1.41
D286A/I287A
site-directed mutagenesis, mutation of a IDH2 residue, highly reduced NAD+ binding, ligand-binding analysis
Saccharomyces cerevisiae
1.1.1.41
H281A
site-directed mutagenesis, mutation of a IDH2 residue, mutant cannot bind NAD+, ligand-binding analysis
Saccharomyces cerevisiae
1.1.1.41
R274A
site-directed mutagenesis, mutation of a IDH1 residue, reduced AMP binding, ligand-binding analysis
Saccharomyces cerevisiae
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.41
additional information
-
additional information
kinetics and ligand binding
Saccharomyces cerevisiae
Localization (protein specific)
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
1.1.1.41
mitochondrion
-
Saccharomyces cerevisiae
5739
-
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
1.1.1.41
Mg2+
binding site at subunit IDH2, active site
Saccharomyces cerevisiae
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.1.1.41
303000
-
sequence calculation
Saccharomyces cerevisiae
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.1.1.41
isocitrate + NAD+
Saccharomyces cerevisiae
enzyme performs the rate-limiting step in the mitochondrial tricarboxylic cycle and is subject to complex allosteric regulation
2-oxoglutarate + CO2 + NADH
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.41
recombinant His-tagged wild-type and mutant enzymes from strain IDH12DELTAL by nickel affinity chromatography
Saccharomyces cerevisiae
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.1.1.41
D-isocitrate + NAD+
catalytic residues binding isocitrate are located on subunit IDH2, while regulatory residues binding isocitrate are located on subunit IDH1
656118
Saccharomyces cerevisiae
2-oxoglutarate + CO2 + NADH
-
-
-
?
1.1.1.41
isocitrate + NAD+
enzyme performs the rate-limiting step in the mitochondrial tricarboxylic cycle and is subject to complex allosteric regulation
656118
Saccharomyces cerevisiae
2-oxoglutarate + CO2 + NADH
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.1.1.41
octamer
alpha4beta4, subunits are termed IDH1 and IDH2
Saccharomyces cerevisiae
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.1.1.41
24
-
assay at
Saccharomyces cerevisiae
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.1.1.41
7.4
-
assay at
Saccharomyces cerevisiae