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Literature summary extracted from

  • Hu, G.; McAlister-Henn, L.
    Novel allosteric properties produced by residue substitutions in the subunit interface of yeast NAD+-specific isocitrate dehydrogenase (2006), Arch. Biochem. Biophys., 453, 207-216.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.41 wild-type and mutant enzymes are expressed in a idh1DELTAidh2DELTA yeast strain which contains deletion/insertion mutations in genomic IDH1 and IDH2 loci Saccharomyces cerevisiae

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.41 I221A residue changes in IDH2 subunits Saccharomyces cerevisiae
1.1.1.41 I221A/V225A/V229A mutant enzyme IDH1-IDH2(I221A/V225A/V229A) shows an about 6fold decrease in apparent affinity for isocitrate measured in the absence or presence of AMP as compared to wild-type enzyme. Mutant enzyme IDH1-IDH2 (I221A/V225A/V229A) exhibits a moderate decrease in apparent affnity for cofactor (about 2.5fold relative to wild-type) in the absence of AMP. The mutant enzyme has acquired the novel property of cooperativity with respect to NAD+, but this cooperativity is not apparent in the presence of AMP. The mutant enzyme with residue changes in only one subunit are active in vivo Saccharomyces cerevisiae
1.1.1.41 S220A residue changes in IDH1 subunits Saccharomyces cerevisiae
1.1.1.41 V214A residue changes in IDH1 subunits Saccharomyces cerevisiae
1.1.1.41 V216A/S220A/V224A IDH1(V216A/S220A/V224A)IDH2 mutant enzyme shows an about 6fold decrease in apparent affinity for isocitrate measured in the absence or presence of AMP as compared to wild-type enzyme. IDH1(V216A/S220A/V224A)IDH2 mutant enzyme exhibits a moderate decrease in apparent affnity for cofactor (about 2.5fold relative to wild-type) in the absence of AMP. The mutant enzyme has acquired the novel property of cooperativity with respect to NAD+, but this cooperativity is not apparent in the presence of AMP. The mutant enzyme with residue changes in only one subunit are active in vivo Saccharomyces cerevisiae
1.1.1.41 V216A/S220A/V224A/I221A/V225A/V229A in the absence of AMP, the IDH1(V216A/S220A/V224A)-IDH2(I221A/V225A/V229A) mutant enzyme demonstrates an about 30fold decrease in apparent Vmax relative to wild-type and a loss of cooperativity with respect to isocitrate and, in the presence of AMP, the apparent affinity of the enzyme for isocitrate is 35fold lower than that of the wild-type enzyme. This mutant enzyme also exhibits a significant decrease in apparent affinity for NAD+ (about 30fold in the absence of AMP and about 10fold in the presence of AMP relative to wild-type). The mutant enzyme has acquired the novel property of cooperativity with respect to NAD+, but this cooperativity is not apparent in the presence of AMP. The mutant enzyme demonstrates decreases in apparent affnity for isocitrate of about 12fold in the absence of AMP and of about 26fold in the presence of AMP relative to wild-type. For this mutant enzyme, with respect to isocitrate, allosteric activation by AMP and cooperativity in the absence of AMP are no longer apparen. The mutant enzyme is not fully functional in vivo Saccharomyces cerevisiae
1.1.1.41 V224A residue changes in IDH1 subunits Saccharomyces cerevisiae
1.1.1.41 V225A residue changes in IDH2 subunits Saccharomyces cerevisiae
1.1.1.41 V229A residue changes in IDH2 subunits Saccharomyces cerevisiae

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.41 37755
-
 4 * 38001, 4 * 37755, the enzyme consists of four IDH1 and four IDH2 subunits, the basic structural unit of the enzyme is an IDH1/IDH2 heterodimer Saccharomyces cerevisiae
1.1.1.41 38001
-
 4 * 38001, 4 * 37755, the enzyme consists of four IDH1 and four IDH2 subunits, the basic structural unit of the enzyme is an IDH1/IDH2 heterodimer Saccharomyces cerevisiae
1.1.1.41 335000
-
 gel filtration Saccharomyces cerevisiae
1.1.1.41 335000
-
 wild-type enzyme, gel filtration Saccharomyces cerevisiae

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.41 Saccharomyces cerevisiae
-
-
-
1.1.1.41 Saccharomyces cerevisiae P28241 IDH2
-
1.1.1.41 Saccharomyces cerevisiae P28834 IDH1
-
1.1.1.41 Saccharomyces cerevisiae MMY011
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.41
-
 Saccharomyces cerevisiae
1.1.1.41 Ni2+-nitrilotriacetate (Ni2+-NTA) column chromatography Saccharomyces cerevisiae

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.41 D-isocitrate + NAD+
-
 Saccharomyces cerevisiae 2-oxoglutarate + CO2 + NADH
-
 ir
1.1.1.41 D-isocitrate + NAD+
-
 Saccharomyces cerevisiae MMY011 2-oxoglutarate + CO2 + NADH
-
 ir
1.1.1.41 isocitrate + NAD+
-
 Saccharomyces cerevisiae 2-oxoglutarate + CO2 + NADH + H+
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.1.41 octamer 4 * 38001, 4 * 37755, the enzyme consists of four IDH1 and four IDH2 subunits, the basic structural unit of the enzyme is an IDH1/IDH2 heterodimer Saccharomyces cerevisiae
1.1.1.41 octamer 4 * IDH1 + 4 * IDH2 Saccharomyces cerevisiae

Synonyms

EC Number Synonyms Comment Organism
1.1.1.41 IDH
-
 Saccharomyces cerevisiae
1.1.1.41 NAD+-specific isocitrate dehydrogenase
-
 Saccharomyces cerevisiae