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Literature summary extracted from

  • Lee, P.; Colman, R.F.
    Implication by site-directed mutagenesis of Arg314 and Tyr316 in the coenzyme site of pig mitochondrial NADP-dependent isocitrate dehydrogenase (2002), Arch. Biochem. Biophys., 401, 81-90.
    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 TB1 as maltose binding fusion proteins Sus scrofa

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.42 K321Q site-directed mutagenesis, kinetics are similar to the wild-type enzyme Sus scrofa
1.1.1.42 R314Q site-directed mutagenesis, 10fold increased Km for NADP+ compared to the wild-type enzyme Sus scrofa
1.1.1.42 R323Q site-directed mutagenesis, kinetics are similar to the wild-type enzyme Sus scrofa
1.1.1.42 Y316F site-directed mutagenesis, kinetics are similar to the wild-type enzyme Sus scrofa
1.1.1.42 Y316L site-directed mutagenesis, 4fold increased Km for NADP+ compared to the wild-type enzyme Sus scrofa

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.42 NAD+
-
 Sus scrofa

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.42 0.0003
-
 Mn2+ pH 7.4, recombinant wild-type enzyme Sus scrofa
1.1.1.42 0.0004
-
 Mn2+ pH 7.4, recombinant mutant Y316F and K323Q Sus scrofa
1.1.1.42 0.0005
-
 Mn2+ pH 7.4, recombinant mutant K321Q Sus scrofa
1.1.1.42 0.0059
-
 isocitrate pH 7.4, recombinant mutant Y316F Sus scrofa
1.1.1.42 0.0062
-
 isocitrate pH 7.4, recombinant mutant K323Q Sus scrofa
1.1.1.42 0.0074
-
 isocitrate pH 7.4, recombinant wild-type enzyme Sus scrofa
1.1.1.42 0.0077
-
 NADP+ pH 7.4, recombinant mutant K323Q Sus scrofa
1.1.1.42 0.008
-
 NADP+ pH 7.4, recombinant mutant Y316F and K321Q Sus scrofa
1.1.1.42 0.009
-
 NADP+ pH 7.4, recombinant wild-type enzyme Sus scrofa
1.1.1.42 0.01
-
 isocitrate pH 7.4, recombinant mutant K321Q Sus scrofa
1.1.1.42 0.032
-
 NADP+ pH 7.4, recombinant mutant Y316L Sus scrofa
1.1.1.42 0.092
-
 NADP+ pH 7.4, recombinant mutant R314Q 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 82000
-
 recombinant mutant Y316F, dynamic light scattering Sus scrofa
1.1.1.42 86000
-
 recombinant wild-type enzyme, dynamic light scattering Sus scrofa
1.1.1.42 87000
-
 recombinant mutants K321Q and R314Q, dynamic light scattering Sus scrofa
1.1.1.42 97900
-
 recombinant mutant Y316L, native PAGE at pH 5.5 Sus scrofa
1.1.1.42 99800
-
 recombinant wild-type enzyme, native PAGE at pH 5.5 Sus scrofa

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 enzymes from Escherichia coli, to homogeneity by amylose affinity chromatography, thrombin digestion, and DEAE ion exchange chromatography 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 7
-
 purified recombinant mutant Y316L Sus scrofa
1.1.1.42 18.1
-
 purified recombinant mutant R314Q Sus scrofa
1.1.1.42 38.6
-
 purified recombinant wild-type enzyme Sus scrofa
1.1.1.42 38.7
-
 purified recombinant mutant K323Q Sus scrofa
1.1.1.42 39.6
-
 purified recombinant mutant K321Q Sus scrofa
1.1.1.42 40.3
-
 purified recombinant mutant Y316F 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+
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.1.42 dimer residues Arg314, Tyr316, Lys321, and Arg323 are not involved in subunit interaction Sus scrofa

Synonyms

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

pH Optimum

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

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.42 NADP+ Arg314 and the aromatic ring of Tyr316 are important for affinity to the cofactor and catalysis, interaction with 2'-phosphate group of NADP+ Sus scrofa

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.1.1.42 8
-
 NAD+ pH 7.4, versus NADPH, recombinant mutant R314Q Sus scrofa
1.1.1.42 11
-
 NAD+ pH 7.4, versus NADPH, recombinant wild-type enzyme and mutant Y316L Sus scrofa
1.1.1.42 12
-
 NAD+ pH 7.4, versus NADPH, recombinant mutant Y316F Sus scrofa