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Literature summary extracted from

  • Kim, T.K.; Lee, P.; Colman, R.F.
    Critical role of Lys212 and Tyr140 in porcine NADP-dependent isocitrate dehydrogenase (2003), J. Biol. Chem., 278, 49323-49331.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.1.1.42 acetic acid mutant Y140T, 106fold activation compared to the wild-type enzyme Sus scrofa
1.1.1.42 ethylamine mutant K212Q, 4fold activation compared to the wild-type enzyme Sus scrofa
1.1.1.42 phenol mutant Y140T, 88fold activation compared to the wild-type enzyme Sus scrofa

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.42 expression of wild-type and mutant enzymes in Escherichia coli Sus scrofa

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.42 K212Q site-directed mutagenesis, highly decreased activity in both reaction directions compared to the wild-type enzyme, altered pH-dependency of the activity Sus scrofa
1.1.1.42 K212R site-directed mutagenesis, highly decreased activity in both reaction directions compared to the wild-type enzyme, altered pH-dependency of the activity Sus scrofa
1.1.1.42 K212Y site-directed mutagenesis, highly decreased activity in both reaction directions compared to the wild-type enzyme, altered pH-dependency of the activity Sus scrofa
1.1.1.42 Y140E site-directed mutagenesis, highly decreased activity in both reaction directions compared to the wild-type enzyme, unaltered Km for isocitrate and NADP+ Sus scrofa
1.1.1.42 Y140F site-directed mutagenesis, highly decreased activity in both reaction directions compared to the wild-type enzyme, unaltered Km for isocitrate and NADP+ Sus scrofa
1.1.1.42 Y140K site-directed mutagenesis, highly decreased activity in both reaction directions compared to the wild-type enzyme, unaltered Km for isocitrate and NADP+ Sus scrofa
1.1.1.42 Y140T site-directed mutagenesis, highly decreased activity in both reaction directions compared to the wild-type enzyme, unaltered Km for isocitrate and NADP+, highly increased activation by added exogenous acetic acid and phenol compared to the wild-type enzyme Sus scrofa

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.42 0.00004
-
 Mn2+ pH 7.4, mutant Y140F Sus scrofa
1.1.1.42 0.000084
-
 Mn2+ pH 7.4, mutant Y140E Sus scrofa
1.1.1.42 0.00011
-
 Mn2+ pH 7.4, wild-type enzyme Sus scrofa
1.1.1.42 0.00042
-
 NADP+ pH 7.4, mutant Y140E Sus scrofa
1.1.1.42 0.00046
-
 NADP+ pH 7.4, mutant Y140T Sus scrofa
1.1.1.42 0.00051
-
 NADP+ pH 7.4, mutant Y140K Sus scrofa
1.1.1.42 0.0006
-
 isocitrate pH 7.4, mutant Y140T Sus scrofa
1.1.1.42 0.0007
-
 isocitrate pH 7.4, mutant Y140E Sus scrofa
1.1.1.42 0.00091
-
 Mn2+ pH 7.4, mutant K212Q Sus scrofa
1.1.1.42 0.0011
-
 Mn2+ pH 7.4, mutant K212Y Sus scrofa
1.1.1.42 0.0023
-
 NADP+ pH 7.4, mutant Y140F Sus scrofa
1.1.1.42 0.0052
-
 isocitrate pH 7.4, mutant Y140F Sus scrofa
1.1.1.42 0.0053
-
 isocitrate pH 7.4, mutant Y140K Sus scrofa
1.1.1.42 0.0073
-
 NADP+ pH 7.4, mutant K212Y Sus scrofa
1.1.1.42 0.0075
-
 isocitrate pH 7.4, wild-type enzyme Sus scrofa
1.1.1.42 0.0092
-
 isocitrate pH 7.4, mutant K212Y Sus scrofa
1.1.1.42 0.0098
-
 isocitrate pH 7.4, mutant K212Q Sus scrofa
1.1.1.42 0.0099
-
 NADP+ pH 7.4, wild-type enzyme Sus scrofa
1.1.1.42 0.014
-
 NADP+ pH 7.4, mutant K212Q Sus scrofa
1.1.1.42 0.018
-
 NADP+ pH 7.4, mutant K212R Sus scrofa
1.1.1.42 0.116
-
 isocitrate pH 7.4, mutant K212R Sus scrofa
1.1.1.42 0.131
-
 Mn2+ pH 7.4, mutant Y140K Sus scrofa
1.1.1.42 0.252
-
 Mn2+ pH 7.4, mutant K212R Sus scrofa

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
1.1.1.42 mitochondrion
-
 Sus scrofa 5739
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.1.42 Mn2+
-
 Sus scrofa

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.42 43000
-
 2 * 43000, recombinant wild-type and mutant enzymes, SDS-PAGE Sus scrofa
1.1.1.42 92000
-
 recombinant wild-type and mutant enzymes, native PAGE Sus scrofa

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.42 Sus scrofa P33198
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.42 recombinant wild-type and mutant enzymes from Escherichia coli, to homogeneity Sus scrofa

Reaction

EC Number Reaction Comment Organism Reaction ID
1.1.1.42 isocitrate + NADP+ = 2-oxoglutarate + CO2 + NADPH + H+ Tyr140 and Lys212 are required for catalytic activity, Tyr140 is the general acid that protonates the substrate after decarboxylation, Lys212 lowers as a positively charged residue the pK of the nearby ionizable group in the enzyme-substrate complex and stabilizes the carbanion formed initially on substrate decarboxylation Sus scrofa
a

Source Tissue

EC Number Source Tissue Comment Organism Textmining
1.1.1.42 heart
-
 Sus scrofa
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.1.42 0.003
-
 purified recombinant mutant K212Y, forward reaction Sus scrofa
1.1.1.42 0.065
-
 purified recombinant mutant K212Q, forward reaction Sus scrofa
1.1.1.42 0.08
-
 purified recombinant mutant Y140E, forward reaction Sus scrofa
1.1.1.42 0.083
-
 purified recombinant mutant Y140T, forward reaction Sus scrofa
1.1.1.42 0.102
-
 purified recombinant mutant Y140F, forward reaction Sus scrofa
1.1.1.42 0.179
-
 purified recombinant mutant Y140K, forward reaction Sus scrofa
1.1.1.42 3.16
-
 purified recombinant mutant K212R, forward reaction Sus scrofa
1.1.1.42 35.1
-
 purified recombinant wild-type enzyme, forward reaction Sus scrofa

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.42 isocitrate + NADP+
-
 Sus scrofa 2-oxoglutarate + CO2 + NADPH + H+
-
 r

Subunits

EC Number Subunits Comment Organism
1.1.1.42 dimer 2 * 43000, recombinant wild-type and mutant enzymes, SDS-PAGE Sus scrofa

Synonyms

EC Number Synonyms Comment Organism
1.1.1.42 NADP-dependent isocitrate dehydrogenase
-
 Sus scrofa

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.42 25
-
 forward reaction, assay at Sus scrofa
1.1.1.42 37
-
 reverse reaction, assay at Sus scrofa

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.42 7.4
-
 assay at Sus scrofa

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.1.1.42 additional information
-
 pH-profile, recombinant wild-type and mutant enzymes Sus scrofa

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.42 NADP+
-
 Sus scrofa
1.1.1.42 NADPH
-
 Sus scrofa