BRENDA - Enzyme Database

Isocitrate binding at two functionally distinct sites in yeast NAD+-specific isocitrate dehydrogenase

Lin, A.P.; McAlister-Henn, L.; J. Biol. Chem. 277, 22475-22483 (2002)

Data extracted from this reference:

Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
1.1.1.41
AMP
allosteric activation
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
A108R/F136Y/T241D/N245D
site-directed mutagenesis, residues of subunit IDH1 mutant is unable to bind AMP, ligand-binding analysis
Saccharomyces cerevisiae
1.1.1.41
A108R/F136Y/T241D/N245D/R114A/Y142F/D248T/D252N
site-directed mutagenesis, residues of subunit IDH1 are A108R, F136Y, T241D, and N245D, residues of subunit IDH2 are R114A, Y142F, D248T, and D252N, mutant is unable to bind AMP, ligand-binding analysis
Saccharomyces cerevisiae
1.1.1.41
R114A/Y142F/D248T/D252N
site-directed mutagenesis, residues of subunit subunit IDH2, ligand-binding analysis
Saccharomyces cerevisiae
1.1.1.41
S92A
site-directed mutagenesis, residue of subunit IDH1, reduction of isocitrate substrate binding sites by half, detrimental effects on isocitrate binding and respective kinetic defects in catalysis and allosteric activation by AMP, ligand-binding analysis
Saccharomyces cerevisiae
1.1.1.41
S92A/S98A
site-directed mutagenesis, residue of subunit IDH1 is S92, residue of subunit IDH2 is S98, ligand-binding analysis
Saccharomyces cerevisiae
1.1.1.41
S98A
site-directed mutagenesis, residue of subunit IDH2, reduction of isocitrate substrate binding sites by half, detrimental effects on isocitrate binding and respective kinetic defects in catalysis and allosteric activation by AMP, 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
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
1.1.1.41
Mg2+
-
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
DL-isocitrate + NAD+
Saccharomyces cerevisiae
-
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
Storage Stability
EC Number
Storage Stability
Organism
1.1.1.41
4C, recombinant wild-type enzyme, in affinity elution buffer containing 50 mM sodium phosphate, pH 7.5, 300 mM NaCl, and 200 mM imidazole, stable for several weeks
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
DL-isocitrate + NAD+
-
656052
Saccharomyces cerevisiae
2-oxoglutarate + CO2 + NADH
-
-
-
?
1.1.1.41
DL-isocitrate + NAD+
isocitrate binds at 2 functionally distinct sites
656052
Saccharomyces cerevisiae
2-oxoglutarate + CO2 + NADH
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
1.1.1.41
More
model of binding sites for isocitrate of the 2 different subunits
Saccharomyces cerevisiae
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
Saccharomyces cerevisiae
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
1.1.1.41
AMP
allosteric activation
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
Saccharomyces cerevisiae
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
1.1.1.41
A108R/F136Y/T241D/N245D
site-directed mutagenesis, residues of subunit IDH1 mutant is unable to bind AMP, ligand-binding analysis
Saccharomyces cerevisiae
1.1.1.41
A108R/F136Y/T241D/N245D/R114A/Y142F/D248T/D252N
site-directed mutagenesis, residues of subunit IDH1 are A108R, F136Y, T241D, and N245D, residues of subunit IDH2 are R114A, Y142F, D248T, and D252N, mutant is unable to bind AMP, ligand-binding analysis
Saccharomyces cerevisiae
1.1.1.41
R114A/Y142F/D248T/D252N
site-directed mutagenesis, residues of subunit subunit IDH2, ligand-binding analysis
Saccharomyces cerevisiae
1.1.1.41
S92A
site-directed mutagenesis, residue of subunit IDH1, reduction of isocitrate substrate binding sites by half, detrimental effects on isocitrate binding and respective kinetic defects in catalysis and allosteric activation by AMP, ligand-binding analysis
Saccharomyces cerevisiae
1.1.1.41
S92A/S98A
site-directed mutagenesis, residue of subunit IDH1 is S92, residue of subunit IDH2 is S98, ligand-binding analysis
Saccharomyces cerevisiae
1.1.1.41
S98A
site-directed mutagenesis, residue of subunit IDH2, reduction of isocitrate substrate binding sites by half, detrimental effects on isocitrate binding and respective kinetic defects in catalysis and allosteric activation by AMP, 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
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
1.1.1.41
Mg2+
-
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
DL-isocitrate + NAD+
Saccharomyces cerevisiae
-
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
Storage Stability (protein specific)
EC Number
Storage Stability
Organism
1.1.1.41
4C, recombinant wild-type enzyme, in affinity elution buffer containing 50 mM sodium phosphate, pH 7.5, 300 mM NaCl, and 200 mM imidazole, stable for several weeks
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
DL-isocitrate + NAD+
-
656052
Saccharomyces cerevisiae
2-oxoglutarate + CO2 + NADH
-
-
-
?
1.1.1.41
DL-isocitrate + NAD+
isocitrate binds at 2 functionally distinct sites
656052
Saccharomyces cerevisiae
2-oxoglutarate + CO2 + NADH
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.1.1.41
More
model of binding sites for isocitrate of the 2 different subunits
Saccharomyces cerevisiae
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