BRENDA - Enzyme Database
show all sequences of 1.1.1.286

Identification of a novel trifunctional homoisocitrate dehydrogenase and modulation of the broad substrate specificity through site-directed mutagenesis

Miyazaki, K.; Biochem. Biophys. Res. Commun. 336, 596-602 (2005)

Data extracted from this reference:

Engineering
Amino acid exchange
Commentary
Organism
additional information
deletion of A80, inability of mutant to use 3-isopropylmalate, isocitrate is 4fold preferred over homoisocitrate
Deinococcus radiodurans
R85V
increased ability to use 3-isopropylmalate
Deinococcus radiodurans
R87T
uses substrate homoisocitrate, but not substrates isocitrate or 3-isopropylmalate
Deinococcus radiodurans
R87V
uses substrate homoisocitrate, but not substrates isocitrate or 3-isopropylmalate
Deinococcus radiodurans
Inhibitors
Inhibitors
Commentary
Organism
Structure
K+
absolute requirement, inhibitory above 0.4 M
Deinococcus radiodurans
Mn2+
inhibitory above 1 mM
Deinococcus radiodurans
NH4+
may substitute for K+, inhibitory above 0.2 M
Deinococcus radiodurans
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.193
-
NAD+
30C, pH 7.8
Deinococcus radiodurans
0.211
-
homoisocitrate
30C, pH 7.8, wild-type
Deinococcus radiodurans
0.291
-
isocitrate
30C, pH 7.8, wild-type
Deinococcus radiodurans
0.465
-
isocitrate
30C, pH 7.8, A80 deletion mutant
Deinococcus radiodurans
0.819
-
homoisocitrate
30C, pH 7.8, A80 deletion mutant
Deinococcus radiodurans
1
-
homoisocitrate
30C, pH 7.8, mutant R87V
Deinococcus radiodurans
1.33
-
3-isopropylmalate
30C, pH 7.8, wild-type
Deinococcus radiodurans
1.5
-
homoisocitrate
30C, pH 7.8, mutant R87T
Deinococcus radiodurans
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
K+
absolute requirement, inhibitory above 0.4 M
Deinococcus radiodurans
Mg2+
divalent cation required, Mg2+ is preferred over Mn2+
Deinococcus radiodurans
Mn2+
divalent cation required, Mg2+ is preferred over Mn2+, inhibitory above 1 mM
Deinococcus radiodurans
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
35727
-
4 * 35751, mass spectrometry, 4 * 35727, calculated
Deinococcus radiodurans
35751
-
4 * 35751, mass spectrometry, 4 * 35727, calculated
Deinococcus radiodurans
154000
-
gel filtration
Deinococcus radiodurans
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Deinococcus radiodurans
-
-
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3-isopropylmalate + NAD+
at 0.1% of the rate with homoisocitrate
660918
Deinococcus radiodurans
?
-
-
-
?
homoisocitrate + NAD+
1.5fold preferred over isocitrate
660918
Deinococcus radiodurans
? + NADH
-
-
-
?
isocitrate + NAD+
-
660918
Deinococcus radiodurans
2-oxoglutarate + CO2 + NADH
-
-
-
?
isocitrate + NADP+
NAD+ is preferred over NADP+
660918
Deinococcus radiodurans
2-oxoglutarate + CO2 + NADPH
-
-
-
?
additional information
no substrate: NADP+
660918
Deinococcus radiodurans
?
-
-
-
-
Subunits
Subunits
Commentary
Organism
tetramer
4 * 35751, mass spectrometry, 4 * 35727, calculated
Deinococcus radiodurans
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.371
-
3-isopropylmalate
30C, pH 7.8, wild-type
Deinococcus radiodurans
5.46
-
homoisocitrate
30C, pH 7.8, A80 deletion mutant
Deinococcus radiodurans
10.9
-
isocitrate
30C, pH 7.8, A80 deletion mutant
Deinococcus radiodurans
20.8
-
homoisocitrate
30C, pH 7.8, mutant R87V
Deinococcus radiodurans
27.8
-
homoisocitrate
30C, pH 7.8, mutant R87T
Deinococcus radiodurans
30.3
-
NAD+
30C, pH 7.8
Deinococcus radiodurans
42.8
-
isocitrate
30C, pH 7.8, wild-type
Deinococcus radiodurans
46.2
-
homoisocitrate
30C, pH 7.8, wild-type
Deinococcus radiodurans
Cofactor
Cofactor
Commentary
Organism
Structure
NAD+
-
Deinococcus radiodurans
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NAD+
-
Deinococcus radiodurans
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
additional information
deletion of A80, inability of mutant to use 3-isopropylmalate, isocitrate is 4fold preferred over homoisocitrate
Deinococcus radiodurans
R85V
increased ability to use 3-isopropylmalate
Deinococcus radiodurans
R87T
uses substrate homoisocitrate, but not substrates isocitrate or 3-isopropylmalate
Deinococcus radiodurans
R87V
uses substrate homoisocitrate, but not substrates isocitrate or 3-isopropylmalate
Deinococcus radiodurans
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
K+
absolute requirement, inhibitory above 0.4 M
Deinococcus radiodurans
Mn2+
inhibitory above 1 mM
Deinococcus radiodurans
NH4+
may substitute for K+, inhibitory above 0.2 M
Deinococcus radiodurans
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.193
-
NAD+
30C, pH 7.8
Deinococcus radiodurans
0.211
-
homoisocitrate
30C, pH 7.8, wild-type
Deinococcus radiodurans
0.291
-
isocitrate
30C, pH 7.8, wild-type
Deinococcus radiodurans
0.465
-
isocitrate
30C, pH 7.8, A80 deletion mutant
Deinococcus radiodurans
0.819
-
homoisocitrate
30C, pH 7.8, A80 deletion mutant
Deinococcus radiodurans
1
-
homoisocitrate
30C, pH 7.8, mutant R87V
Deinococcus radiodurans
1.33
-
3-isopropylmalate
30C, pH 7.8, wild-type
Deinococcus radiodurans
1.5
-
homoisocitrate
30C, pH 7.8, mutant R87T
Deinococcus radiodurans
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
K+
absolute requirement, inhibitory above 0.4 M
Deinococcus radiodurans
Mg2+
divalent cation required, Mg2+ is preferred over Mn2+
Deinococcus radiodurans
Mn2+
divalent cation required, Mg2+ is preferred over Mn2+, inhibitory above 1 mM
Deinococcus radiodurans
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
35727
-
4 * 35751, mass spectrometry, 4 * 35727, calculated
Deinococcus radiodurans
35751
-
4 * 35751, mass spectrometry, 4 * 35727, calculated
Deinococcus radiodurans
154000
-
gel filtration
Deinococcus radiodurans
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3-isopropylmalate + NAD+
at 0.1% of the rate with homoisocitrate
660918
Deinococcus radiodurans
?
-
-
-
?
homoisocitrate + NAD+
1.5fold preferred over isocitrate
660918
Deinococcus radiodurans
? + NADH
-
-
-
?
isocitrate + NAD+
-
660918
Deinococcus radiodurans
2-oxoglutarate + CO2 + NADH
-
-
-
?
isocitrate + NADP+
NAD+ is preferred over NADP+
660918
Deinococcus radiodurans
2-oxoglutarate + CO2 + NADPH
-
-
-
?
additional information
no substrate: NADP+
660918
Deinococcus radiodurans
?
-
-
-
-
Subunits (protein specific)
Subunits
Commentary
Organism
tetramer
4 * 35751, mass spectrometry, 4 * 35727, calculated
Deinococcus radiodurans
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.371
-
3-isopropylmalate
30C, pH 7.8, wild-type
Deinococcus radiodurans
5.46
-
homoisocitrate
30C, pH 7.8, A80 deletion mutant
Deinococcus radiodurans
10.9
-
isocitrate
30C, pH 7.8, A80 deletion mutant
Deinococcus radiodurans
20.8
-
homoisocitrate
30C, pH 7.8, mutant R87V
Deinococcus radiodurans
27.8
-
homoisocitrate
30C, pH 7.8, mutant R87T
Deinococcus radiodurans
30.3
-
NAD+
30C, pH 7.8
Deinococcus radiodurans
42.8
-
isocitrate
30C, pH 7.8, wild-type
Deinococcus radiodurans
46.2
-
homoisocitrate
30C, pH 7.8, wild-type
Deinococcus radiodurans
Other publictions for EC 1.1.1.286
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [C]
Temperature Range [C]
Temperature Stability [C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [C] (protein specific)
Temperature Range [C] (protein specific)
Temperature Stability [C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
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Shimizu
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1
1
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16
1
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1
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1
1
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32
-
1
-
4
-
-
-
1
-
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-
8
1
1
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16
1
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3
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2
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1
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6
1
1
-
-
2
1
-
-
2
-
-
-
-
-
1
2
-
-
-
-
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2
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6
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1
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6
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1
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1
1
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2
2
740000
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-
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-
8
1
1
-
-
2
1
-
-
2
-
-
-
-
-
1
2
1
-
-
-
1
-
2
-
1
-
6
-
-
-
1
-
-
-
-
8
1
1
-
-
2
1
-
-
-
-
3
3
-
2
2
685232
Lin
Chemical mechanism of homoisoc ...
Saccharomyces cerevisiae
Biochemistry
47
4169-4180
2008
-
-
-
-
-
-
3
1
-
-
-
1
-
1
-
-
-
2
-
-
-
-
3
-
1
-
-
-
1
1
-
1
1
-
-
-
-
-
1
-
-
-
-
3
1
1
-
-
-
1
-
-
-
-
-
-
-
-
3
-
1
-
-
-
1
1
-
-
-
-
-
-
-
-
696220
Lin
Potassium is an activator of h ...
Saccharomyces cerevisiae
Biochemistry
47
10809-10815
2008
-
-
-
-
-
-
4
-
-
4
-
1
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
1
3
-
-
-
-
-
1
-
-
-
-
4
3
-
-
4
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
660907
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331
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-
-
1
-
-
-
-
3
-
-
-
-
-
3
-
-
1
-
-
-
-
-
4
-
-
-
-
3
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
3
-
-
-
-
-
-
-
1
-
-
-
-
4
-
-
-
-
3
-
-
-
-
-
-
-
-
-
-
660918
Miyazaki
Identification of a novel trif ...
Deinococcus radiodurans
Biochem. Biophys. Res. Commun.
336
596-602
2005
-
-
-
-
4
-
3
8
-
3
3
-
-
1
-
-
-
-
-
-
-
-
5
1
-
-
-
8
-
-
-
1
-
-
-
-
-
-
1
-
4
-
-
3
-
8
-
3
3
-
-
-
-
-
-
-
-
-
5
1
-
-
-
8
-
-
-
-
-
-
-
-
-
-
656127
Miyazaki
Characterization of homoisocit ...
Thermus thermophilus, Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039
J. Biol. Chem.
278
1864-1871
2003
-
-
1
-
1
-
-
2
-
-
-
-
-
2
-
-
-
-
-
-
-
-
6
1
-
-
1
2
-
-
-
-
-
-
-
-
-
1
-
-
1
-
-
-
-
2
-
-
-
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-
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-
-
-
-
-
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6
1
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-
1
2
-
-
-
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-
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-
-
-