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Literature summary extracted from

  • Miyazaki, K.
    Identification of a novel trifunctional homoisocitrate dehydrogenase and modulation of the broad substrate specificity through site-directed mutagenesis (2005), Biochem. Biophys. Res. Commun., 336, 596-602.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.87 DNA and amino acid sequence determination and analysis, overexpression in Escherichia coli strain OM17 Deinococcus radiodurans

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.87 A80del site-directed mutagenesis, the mutant shows altered substrate specificity preferring isocitrate to homoisocitrate, it is unable to oxidize 3-isopropylmalate, the specificity is similar to the enzyme from Thermus thermophilus Deinococcus radiodurans
1.1.1.87 additional information modulation of the broad substrate specificity of the trifunctional enzyme through site-directed mutagenesis, overview Deinococcus radiodurans
1.1.1.87 R87T site-directed mutagenesis, the mutant oxidizes homoisocitrate, but not isocitrate and 3-isopropylmalate Deinococcus radiodurans
1.1.1.87 R87V site-directed mutagenesis, the mutant oxidizes homoisocitrate, but not isocitrate and 3-isopropylmalate Deinococcus radiodurans
1.1.1.286 additional information deletion of A80, inability of mutant to use 3-isopropylmalate, isocitrate is 4fold preferred over homoisocitrate Deinococcus radiodurans
1.1.1.286 R85V increased ability to use 3-isopropylmalate Deinococcus radiodurans
1.1.1.286 R87T uses substrate homoisocitrate, but not substrates isocitrate or 3-isopropylmalate Deinococcus radiodurans
1.1.1.286 R87V uses substrate homoisocitrate, but not substrates isocitrate or 3-isopropylmalate Deinococcus radiodurans

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.286 K+ absolute requirement, inhibitory above 0.4 M Deinococcus radiodurans
1.1.1.286 Mn2+ inhibitory above 1 mM Deinococcus radiodurans
1.1.1.286 NH4+ may substitute for K+, inhibitory above 0.2 M Deinococcus radiodurans

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.87 0.211
-
 homoisocitrate pH 7.8, 25°C, recombinant wild-type enzyme Deinococcus radiodurans
1.1.1.87 0.291
-
 isocitrate pH 7.8, 25°C, recombinant wild-type enzyme Deinococcus radiodurans
1.1.1.87 0.465
-
 isocitrate pH 7.8, 25°C, recombinant mutant A80del Deinococcus radiodurans
1.1.1.87 0.819
-
 homoisocitrate pH 7.8, 25°C, recombinant mutant A80del Deinococcus radiodurans
1.1.1.87 1
-
 homoisocitrate pH 7.8, 25°C, recombinant mutant R87V Deinococcus radiodurans
1.1.1.87 1.33
-
 3-isopropylmalate pH 7.8, 25°C, recombinant wild-type enzyme Deinococcus radiodurans
1.1.1.87 1.5
-
 homoisocitrate pH 7.8, 25°C, recombinant mutant R87T Deinococcus radiodurans
1.1.1.286 0.193
-
 NAD+ 30°C, pH 7.8 Deinococcus radiodurans
1.1.1.286 0.211
-
 homoisocitrate 30°C, pH 7.8, wild-type Deinococcus radiodurans
1.1.1.286 0.291
-
 isocitrate 30°C, pH 7.8, wild-type Deinococcus radiodurans
1.1.1.286 0.465
-
 isocitrate 30°C, pH 7.8, A80 deletion mutant Deinococcus radiodurans
1.1.1.286 0.819
-
 homoisocitrate 30°C, pH 7.8, A80 deletion mutant Deinococcus radiodurans
1.1.1.286 1
-
 homoisocitrate 30°C, pH 7.8, mutant R87V Deinococcus radiodurans
1.1.1.286 1.33
-
 3-isopropylmalate 30°C, pH 7.8, wild-type Deinococcus radiodurans
1.1.1.286 1.5
-
 homoisocitrate 30°C, pH 7.8, mutant R87T Deinococcus radiodurans

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.1.87 K+ required Deinococcus radiodurans
1.1.1.87 Mn2+ required Deinococcus radiodurans
1.1.1.286 K+ absolute requirement, inhibitory above 0.4 M Deinococcus radiodurans
1.1.1.286 Mg2+ divalent cation required, Mg2+ is preferred over Mn2+ Deinococcus radiodurans
1.1.1.286 Mn2+ divalent cation required, Mg2+ is preferred over Mn2+, inhibitory above 1 mM Deinococcus radiodurans

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.87 40000
-
 4 * 40000, recombinant enzyme, SDS-PAGE Deinococcus radiodurans
1.1.1.87 154000
-
 recombinant enzyme, gel filtration Deinococcus radiodurans
1.1.1.286 35727
-
 4 * 35751, mass spectrometry, 4 * 35727, calculated Deinococcus radiodurans
1.1.1.286 35751
-
 4 * 35751, mass spectrometry, 4 * 35727, calculated Deinococcus radiodurans
1.1.1.286 154000
-
 gel filtration Deinococcus radiodurans

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.87 Deinococcus radiodurans
-
-
-
1.1.1.286 Deinococcus radiodurans
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.87 recombinant enzyme from Escherichia coli strain OM17 by anion exchange and hydrophobic interaction chromatography, followed by another step of anion exchange chromatography Deinococcus radiodurans

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.1.87 additional information
-
-
 Deinococcus radiodurans

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.87 3-isopropylmalate + NAD+
-
 Deinococcus radiodurans ?
-
 ?
1.1.1.87 homoisocitrate + NAD+ 1.5fold preferred to isocitrate Deinococcus radiodurans ?
-
 ?
1.1.1.87 isocitrate + NAD+
-
 Deinococcus radiodurans ?
-
 ?
1.1.1.286 3-isopropylmalate + NAD+ at 0.1% of the rate with homoisocitrate Deinococcus radiodurans ?
-
 ?
1.1.1.286 homoisocitrate + NAD+ 1.5fold preferred over isocitrate Deinococcus radiodurans ? + NADH
-
 ?
1.1.1.286 isocitrate + NAD+
-
 Deinococcus radiodurans 2-oxoglutarate + CO2 + NADH + H+
-
 ?
1.1.1.286 isocitrate + NADP+ NAD+ is preferred over NADP+ Deinococcus radiodurans 2-oxoglutarate + CO2 + NADPH + H+
-
 ?
1.1.1.286 additional information no substrate: NADP+ Deinococcus radiodurans ?
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.1.87 tetramer 4 * 40000, recombinant enzyme, SDS-PAGE Deinococcus radiodurans
1.1.1.286 tetramer 4 * 35751, mass spectrometry, 4 * 35727, calculated Deinococcus radiodurans

Synonyms

EC Number Synonyms Comment Organism
1.1.1.87 HICDH
-
 Deinococcus radiodurans

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.87 25
-
 assay at Deinococcus radiodurans

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.87 0.371
-
 3-isopropylmalate pH 7.8, 25°C, recombinant wild-type enzyme Deinococcus radiodurans
1.1.1.87 5.46
-
 homoisocitrate pH 7.8, 25°C, recombinant mutant A80del Deinococcus radiodurans
1.1.1.87 10.9
-
 isocitrate pH 7.8, 25°C, recombinant mutant A80del Deinococcus radiodurans
1.1.1.87 20.8
-
 homoisocitrate pH 7.8, 25°C, recombinant mutant R87V Deinococcus radiodurans
1.1.1.87 27.8
-
 homoisocitrate pH 7.8, 25°C, recombinant mutant R87T Deinococcus radiodurans
1.1.1.87 42.8
-
 isocitrate pH 7.8, 25°C, recombinant wild-type enzyme Deinococcus radiodurans
1.1.1.87 46.2
-
 homoisocitrate pH 7.8, 25°C, recombinant wild-type enzyme Deinococcus radiodurans
1.1.1.286 0.371
-
 3-isopropylmalate 30°C, pH 7.8, wild-type Deinococcus radiodurans
1.1.1.286 5.46
-
 homoisocitrate 30°C, pH 7.8, A80 deletion mutant Deinococcus radiodurans
1.1.1.286 10.9
-
 isocitrate 30°C, pH 7.8, A80 deletion mutant Deinococcus radiodurans
1.1.1.286 20.8
-
 homoisocitrate 30°C, pH 7.8, mutant R87V Deinococcus radiodurans
1.1.1.286 27.8
-
 homoisocitrate 30°C, pH 7.8, mutant R87T Deinococcus radiodurans
1.1.1.286 30.3
-
 NAD+ 30°C, pH 7.8 Deinococcus radiodurans
1.1.1.286 42.8
-
 isocitrate 30°C, pH 7.8, wild-type Deinococcus radiodurans
1.1.1.286 46.2
-
 homoisocitrate 30°C, pH 7.8, wild-type Deinococcus radiodurans

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.87 7.8
-
 assay at Deinococcus radiodurans

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.87 NAD+
-
 Deinococcus radiodurans
1.1.1.286 NAD+
-
 Deinococcus radiodurans