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Literature summary extracted from

  • Huang, Y.C.; Colman, R.F.
    Evaluation by mutagenesis of the roles of His309, His315, and His319 in the coenzyme site of pig heart NADP-dependent isocitrate dehydrogenase (2002), Biochemistry, 41, 5637-5643.
    View publication on PubMed

Cloned(Commentary)

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

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.42 H309F site-directed mutagenesis, inactive mutant, poor cofactor binding, altered secondary structure Sus scrofa
1.1.1.42 H309Q site-directed mutagenesis, inactive mutant, poor cofactor binding, altered secondary structure Sus scrofa
1.1.1.42 H315Q site-directed mutagenesis, 40fold increased Km for NADP+ compared to the wild-type enzyme Sus scrofa
1.1.1.42 H319Q site-directed mutagenesis, cofactor binding and kinetics similar to the wild-type enzyme, slightly reduced activity Sus scrofa

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.42 NADPH
-
 Sus scrofa

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.42 additional information
-
 additional information measurement methods, the maltose binding fusion protein of the recombinant enzymes alters the kinetic parameters, overview Sus scrofa
1.1.1.42 0.00033
-
 Mn2+ pH 7.4, 25°C, recombinant wild-type Sus scrofa
1.1.1.42 0.00039
-
 Mn2+ pH 7.4, 25°C, recombinant mutant H319Q Sus scrofa
1.1.1.42 0.0027
-
 Mn2+ pH 7.4, 25°C, recombinant mutant H315Q Sus scrofa
1.1.1.42 0.0051
-
 NADP+ pH 7.4, 25°C, recombinant mutant H319Q Sus scrofa
1.1.1.42 0.0056
-
 NADP+ pH 7.4, 25°C, recombinant wild-type Sus scrofa
1.1.1.42 0.0083
-
 DL-isocitrate pH 7.4, 25°C, recombinant mutant H319Q Sus scrofa
1.1.1.42 0.0084
-
 DL-isocitrate pH 7.4, 25°C, recombinant wild-type Sus scrofa
1.1.1.42 0.0087
-
 DL-isocitrate pH 7.4, 25°C, recombinant mutant H315Q Sus scrofa
1.1.1.42 0.218
-
 NADP+ pH 7.4, 25°C, recombinant mutant H315Q 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+ required, not bound normally Sus scrofa

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.42 46600
-
 2 * 46600, recombinant thrombin cleaved wild-type and mutant enzymes, SDS-PAGE Sus scrofa
1.1.1.42 90000
-
 recombinant maltose binding fusion proteins, wild-type and mutant enzymes, gel filtration Sus scrofa

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.42 DL-isocitrate + NADP+ Sus scrofa
-
 2-oxoglutarate + CO2 + NADPH
-
 ?

Organism

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

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.42 recombinant wild-type and mutant maltose binding fusion proteins from Escherichia coli by amylose affinity and size exclusion chromatography Sus scrofa

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 4.1
-
 purified recombinant mutant H315Q Sus scrofa
1.1.1.42 20.1
-
 purified recombinant mutant H319Q Sus scrofa
1.1.1.42 37.8
-
 purified recombinant wild-type enzyme Sus scrofa

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.42 DL-isocitrate + NADP+
-
 Sus scrofa 2-oxoglutarate + CO2 + NADPH
-
 ?
1.1.1.42 Ds-isocitrate + NADP+
-
 Sus scrofa 2-oxoglutarate + CO2 + NADPH
-
 ?
1.1.1.42 additional information His319 and His315 are not responsible for enzyme Fe2+-isocitrate cleavage Sus scrofa ?
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.1.42 dimer 2 * 46600, recombinant thrombin cleaved 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
-
 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 5 8
-
 Sus scrofa

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.42 NADP+ binding of 1 mol NADP+ per enzyme subunit, His309 is highly important for cofactor binding, HIs315 is involved, while His319 is not Sus scrofa