Information on EC 1.1.1.41 - isocitrate dehydrogenase (NAD+)

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea

EC NUMBER
COMMENTARY hide
1.1.1.41
-
RECOMMENDED NAME
GeneOntology No.
isocitrate dehydrogenase (NAD+)
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
isocitrate + NAD+ = 2-oxoglutarate + CO2 + NADH
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
-
-
-
-
oxidative decarboxylation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
reductive carboxylation
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Biosynthesis of antibiotics
-
-
Biosynthesis of secondary metabolites
-
-
Citrate cycle (TCA cycle)
-
-
citric acid cycle
-
-
L-glutamine biosynthesis III
-
-
Metabolic pathways
-
-
Microbial metabolism in diverse environments
-
-
TCA cycle II (plants and fungi)
-
-
TCA cycle III (animals)
-
-
SYSTEMATIC NAME
IUBMB Comments
isocitrate:NAD+ oxidoreductase (decarboxylating)
Requires Mn2+ or Mg2+ for activity. Unlike EC 1.1.1.42, isocitrate dehydrogenase (NADP+), oxalosuccinate cannot be used as a substrate. In eukaryotes, isocitrate dehydrogenase exists in two forms: an NAD+-linked enzyme found only in mitochondria and displaying allosteric properties, and a non-allosteric, NADP+-linked enzyme that is found in both mitochondria and cytoplasm [7]. The enzyme from some species can also use NADP+ but much more slowly [9].
CAS REGISTRY NUMBER
COMMENTARY hide
9001-58-5
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain ON107
SwissProt
Manually annotated by BRENDA team
Landsberg
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
pigeon
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
strain H37Rv
-
-
Manually annotated by BRENDA team
strain H37Rv
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
W 6
-
-
Manually annotated by BRENDA team
W 6
-
-
Manually annotated by BRENDA team
Rhodosporidium toruloides
Rhodosporidium toruloides AS 2.1389
strain MMY011
-
-
Manually annotated by BRENDA team
Sarcophaga barbata
-
-
-
Manually annotated by BRENDA team
gene SlIDH1
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
waterbug
-
-
-
Manually annotated by BRENDA team
citrate-overproducing yeast, strain BKM Y-2373
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
ssp. mobiliss
-
-
Manually annotated by BRENDA team
ssp. mobiliss
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
metabolism
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
8-(4-bromo-2,3-dioxobutylthio)-NAD+ + isocitrate
2-oxoglutarate + CO2 + 8-(4-bromo-2,3-dioxobutylthio)-NADH
show the reaction diagram
-
-
-
-
?
D-homoisocitrate + NAD+
? + NADH
show the reaction diagram
-
-
-
-
?
D-isocitrate + NAD+
2-oxoglutarate + CO2 + NADH
show the reaction diagram
DL-isocitrate + NAD+
2-oxoglutarate + CO2 + NADH
show the reaction diagram
isocitrate + NAD+
2-oxoglutarate + CO2 + NADH
show the reaction diagram
isocitrate + NAD+
2-oxoglutarate + CO2 + NADH + H+
show the reaction diagram
isocitrate + NAD+
2-oxoglutarate + NADH + H+ + CO2
show the reaction diagram
isocitrate + NADP+
2-oxoglutarate + CO2 + NADPH
show the reaction diagram
threo-Ds-isocitrate + NAD+
2-oxoglutarate + CO2 + NADH
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
D-isocitrate + NAD+
2-oxoglutarate + CO2 + NADH
show the reaction diagram
-
-
-
-
?
DL-isocitrate + NAD+
2-oxoglutarate + CO2 + NADH
show the reaction diagram
isocitrate + NAD+
2-oxoglutarate + CO2 + NADH
show the reaction diagram
isocitrate + NAD+
2-oxoglutarate + NADH + H+ + CO2
show the reaction diagram
threo-Ds-isocitrate + NAD+
2-oxoglutarate + CO2 + NADH
show the reaction diagram
-
-
-
-
?
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
-
the enzyme is also active with NADP+ and NADPH, cf. EC 1.1.1.42
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Zn2+
-
less effective than Mn2+ or Mg2+
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
130-nucleotide transcript containing the 5'-untranslated regions of the yeast mitochondrial COX2 mRNA
-
inhibition with a 50% reduction in activity observed with 40 nM mRNA
-
2-hydroxyethylamine-N,N-diacetate
-
-
2-oxoglutarate
2-Oxopropane-1,1,3-tricarboxylate
-
only in presence of ADP
beta-mercapto-alpha-ketoglutarate
-
inhibition in presence and absence of AMP, competitive inhibition
bilirubin
-
-
Chicago sky blue
-
-
citrate
Congo red
-
-
D,L-hibiscusate
-
with constant concentrations of isocitrate, NAD+ and Mg2+ up to 10 mM and without AMP, D,L-hibiscusate is an activator
D,L-threo-alpha-methylisocitrate
-
inhibition in presence and absence of AMP, competitive inhibition
D-malate
-
inhibition only in absence of AMP
D-Tartrate
-
inhibition only in absence of AMP
Diamide
EDTA
-
-
EGTA
-
-
fluorescein
-
halogenated derivatives
Fluorocitrate
-
with constant concentrations of isocitrate, NAD+ and Mg2+ up to 10 mM and without AMP, fluorocitrate is an activator
fumarate
-
inhibition only in absence of AMP
gamma-aminobutyric acid
-
slight inhibition
glutamate
-
slight inhibition
glyoxylate
homocitrate
-
inhibition only in absence of AMP
L-garciniate
-
inhibition in presence and absence of AMP
-
L-glutamate
Rhodosporidium toruloides
-
-
L-hibiscusate
-
with constant concentrations of isocitrate, NAD+ and Mg2+ up to 10 mM and without AMP, L-hibiscusate is an activator
-
L-Malate
-
inhibition only in absence of AMP, competitive inhibition
L-Tartrate
-
inhibition only in absence of AMP
Maleate
-
inhibition only in absence of AMP
malonate
-
inhibition only in absence of AMP
Mg2+
-
free form
mitochondrial COX2 UTR RNA
-
-
-
molybdate
mRNA
-
enzyme specifically binds to 5'-untranslated regions of yeast mitochondrial mRNAs, and transcripts containing these regions allosterically inhibit the enzyme, which can be relieved by activator AMP in presence of isocitrate, complex formation, overview
nitrilotriacetate
-
-
oxaloacetate
p-hydroxymercuribenzoate
-
-
pyruvate
-
with oxaloacetate, concomitantly added, less effective than glyoxylate + oxaloacetate
succinate
Succinic semialdehyde
Sulfobromophthalein
-
-
Thiocyanate
-
-
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
DTT
-
; decreases disulfide content of the enzyme, kinetics of enzyme mutants in presence or absence of DTT, overview
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.03 - 0.45
D,L-isocitrate
0.09 - 62.2
D-isocitrate
0.075 - 0.385
DL-isocitrate
0.0183
homoisocitrate
-
6fold His-tagged enzyme, at 70C in 50 mM N-2-hydroxyethylpiperadine-N0-2-ethanesulfonate-NaOH (pH 7.8), 0.2 M KCl, 0.2 mM MnCl2, 1mM NAD+
0.0164 - 41
isocitrate
0.22 - 0.39
Mn2+
0.004 - 4.7
NAD+
0.017 - 8.2
NADP+
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.27 - 73
D-isocitrate
1.5 - 124
DL-isocitrate
13.7
homoisocitrate
Pyrococcus horikoshii
-
6fold His-tagged enzyme, at 70C in 50 mM N-2-hydroxyethylpiperadine-N0-2-ethanesulfonate-NaOH (pH 7.8), 0.2 M KCl, 0.2 mM MnCl2, 1 mM NAD+
0.27 - 36.8
isocitrate
2.49 - 112
NAD+
14 - 168
NADP+
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.000004 - 0.00165
DL-isocitrate
0.000002 - 1567
NAD+
0.000003 - 821
NADP+
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.063 - 0.18
NADH
0.26
NADPH
-
pH 7.5
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.084
-
mitochondria from green leaves, forward reaction
0.092
-
partially purified enzyme, idh-II mutant
0.096
-
mitochondria from etiolated leaves, forward reaction
0.15
-
enzyme extract of cells in exponential growth phase
0.16
-
enzyme extract of cells in stationary growth phase
0.169
-
partially purified enzyme, wild-type
0.17
-
enzyme extract of cells in growth phase with acid synthesis
0.2
-
D234C, alpha-subunit mutant, in 20 mM D-isocitrate and 1 mM MnSO4
1.4
-
D230C, alpha-subunit mutant, in 20 mM D-isocitrate and 1 mM MnSO4
2.2
-
purified recombinant mutant gamma-R97Q
2.9
-
-
3.7
-
D215N, gamma-subunit mutant, in 20 mM D-isocitrate and 1 mM MnSO4
13.5
-
purified recombinant mutant beta-R99Q
21.7
-
purified recombinant wild-type enzyme
27 - 31
-
-
29
-
D206N, alpha-subunit mutant, in 20 mM D-isocitrate and 1 mM MnSO4
50.5
-
with 0.12 mM NADP+ and 2 mM D-isocitrate
75.5
-
with 0.12 mM NADP+ and 2 mM D-isocitrate
120
purified native enzyme
additional information
-
no activity on 3-isopropylmalate
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5
-
-
7 - 7.2
-
-
7.2
-
assay at
7.2 - 8.2
Rhodosporidium toruloides
-
-
8.1 - 8.3
-
-
8.1
-
vertebrate muscle
8.4
-
-
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 8.6
-
-
6 - 8.5
-
-
6 - 8
-
-
6.5 - 8.5
6.5 - 10
-
activity range, profile overview
7.2 - 9.2
-
activity range, profile overview
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
70
-
assay at
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
12 - 37
-
-
25 - 55
-
activity range, profile overview
25 - 60
-
and above, activity range, profile overview
80 - 95
-
activity at 80C is about 50% compared to the activity at 90C
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
malignant
Manually annotated by BRENDA team
-
multiple
Manually annotated by BRENDA team
At4g35650 is not expressed in vegetative organs but is mainly expressed in the pollen
Manually annotated by BRENDA team
apex; apex; apex; apex
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
32850
IDH IV, without the predicted mitochondrial target peptide, calculated from sequence of amino acids
35910
IDH III, without the predicted mitochondrial target peptide, calculated from sequence of amino acids
36040
IDH VI, without the predicted mitochondrial target peptide, calculated from sequence of amino acids
36200
IDH V, without the predicted mitochondrial target peptide, calculated from sequence of amino acids
36810
IDH II, without the predicted mitochondrial target peptide, calculated from sequence of amino acids
37500
-
IDH2, SDS-PAGE
38900
IDH I, without the predicted mitochondrial target peptide, calculated from sequence of amino acids
39590
IDH II, with the predicted mitochondrial target peptide, calculated from sequence of amino acids
39630
IDH I, with the predicted mitochondrial target peptide, calculated from sequence of amino acids
39960
IDH III, with the predicted mitochondrial target peptide, calculated from sequence of amino acids
40580
IDH VI, with the predicted mitochondrial target peptide, calculated from sequence of amino acids
40620
IDH V, with the predicted mitochondrial target peptide, calculated from sequence of amino acids
45000
-
IDHa, SDS-PAGE
74000
-
recombinant His6-tagged enzyme, gel filtration
90000
gel filtration and native PAGE
123000
-
disulfide bonded form of subunit IDH2, nonreducing PAGE
145000
-
gel filtration
191800
Rhodosporidium toruloides
-
gel filtration
224000
-
ultracentrifugation
238000
-
gel filtration
245000
-
sedimentation equilibrium analysis
250000
-
Superose 6 gel filtration
260000
-
dynamic light scattering
290000
-
Superdex 200 gel filtration
300000 - 340000
-
gel filtration
303000
-
sequence calculation
315000
-
about, recombinant wild-type and mutant enzymes, gel filtration
320000
335000
gel filtration; wild-type enzyme, gel filtration; wild-type enzyme, gel filtration
338000
-
gel filtration
412000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
heterotetramer
homodimer
monomer
octamer
tetramer
trimer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
-
-
phosphoprotein
-
catalytically important phosphorylation site at Ser102
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant isocitrate dehydrogenase complexed with NAD+ and citrate solved to a resolution of 1.9 A, hanging drop vapor diffusion method
-
crystallization in artificial mother liquor supplemented with 100 mM NAD+
-
sitting drop vapour diffusion method, using 3-5% (w/v) PEG 20000, 0.1 M MES pH 5.7 and 0.2 M CaCl2
-
hanging drop vapor diffusion method
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 8.5
-
-
286757
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0
Rhodosporidium toruloides
-
full loss in activity in a few hours
42
-
10 min, 50% loss in activity
50
-
purified recombinant enzyme, inactivation
102
-
the apparent melting temperature is 102.3C, mutant enzyme D328K
103
-
the apparent melting temperature is 102.9C, mutant enzyme D328K/I329Y
additional information
-
thermal denaturation is an irreversible process
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
20 h in medium containing ADP
20% glycerol stabilizes during storage
-
ICDH-2 is more stable to urea than ICDH-1
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-80C, partially purified enzyme, several months, stable
-
-90C, no loss of activity after 6 weeks
4C, glycerol 20%, 35-40% of original activity after 96 h, membrane-associated ICDH
-
4C, recombinant wild-type enzyme, in affinity elution buffer containing 50 mM sodium phosphate, pH 7.5, 300 mM NaCl, and 200 mM imidazole, stable for several weeks
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
129fold to homogeneity by ammonium sulfate fractionation, hydrophobic chromatography on an octyl resin, and gel filtration
-
ammonium sulfate fractionation and gel filtration
-
gel filtration
-
Hi-Trap chelating column chromatography and Superdex 200 gel filtration
-
Hi-Trap chelating HP column chromatography
-
mitochondrial preparation
-
multi-step purification of the native enzyme, 520fold to homogeneity, partial purification of the recombinant enzyme from Escherichia coli
Ni-NTA column chromatography
-
Ni2+-affinity chromatography
Ni2+-nitrilotriacetate (Ni2+-NTA) column chromatography
Ni2+-nitrilotriacetic acid resin chromatography
-
Ni2+-nitrilotriacetic acid superflow resin chromatography
-
partially by mitochondrion isolation in a Percoll gradient, and ammonium sulfate precipitation in the 60-80% fraction, DEAE cellulose chromatography, and desalting
-
recombinant His-tagged enzyme from Escherichia coli strain BL21-Gold(DE3) by nickel affinity chromatography
-
recombinant His-tagged wild-type and mutant enzymes from strain IDH12DELTAL by nickel affinity chromatography
-
recombinant His6-tagged enzyme from Escherichia coli strain BL21 (DE3)
-
recombinant wild-type and mutant enzymes from Escherichia coli
recombinant wild-type and mutant NAD-IDHs from Escherichia coli by ammonium sulfate fractionation, anion exchange chromatography, ultrafiltration, and gel filtration
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cloned and overexpressed in Escherichia coli
-
DNA and amino acid sequence determination and analysis, genotyping and sequence comparison with the acient ancestor teosinte, Zea mays ssp. Parviglumis, overview
-
expressed in Escherichia coli
expressed in Escherichia coli BL21(DE3)RP cells
-
expressed in Escherichia coli BL21-Gold (DE3) cells
-
expressed in Escherichia coli strain BL21-CodonPlus (DE3)-RIL
-
expressed in Saccharomyces cerevisiae
expression in Escherichia coli
-
expression of His-tagged wild-type and mutant enzymes in yeast strain IDH12DELTAL deficient in both subunits of the enzyme
-
expression of wild-type and mutant NAD-IDHs in Escherichia coli
-
gene icd, DNA and amino acid sequence determination and analysis, expression in enzyme-deficient Escherichia coli strain EB106
gene idh, DNA and amino acid sequence determination and analysis, sequence comparisons, expression of His6-tagged enzyme in Escherichia coli strain BL21 (DE3)
-
gene idh, expression of wild-type and mutant enzymes in Escherichia coli glutamate auxotrophic strain Dicd::kanr fused to gene icd, the endogenous icd gene is replaced by the kanamycin resistance gene, the wild-type phenotype of the icd defective strain on minimal medium without glutamate is restored
-
gene RtIDH1, DNA and amino acid sequence determination and analysis, sequence comparison, expression in the Saccharomyces cerevisiae strain BY4741 idhDELTA mutant, RtIdh expression leads to increased intracellular lipid content and extracellular citrate concentration with increasing carbon/nitrogen molar ratio of the media under nitrogen limiting conditions; gene RtIDH2, DNA and amino acid sequence determination and analysis, sequence comparison, expression in the Saccharomyces cerevisiae strain BY4741 idhDELTA mutant, RtIdh expression leads to increased intracellular lipid content and extracellular citrate concentration with increasing carbon/nitrogen molar ratio of the media under nitrogen limiting conditions
Rhodosporidium toruloides
gene SlIDH1, DNA and amino acid sequence determination and analysis, phylogenetic analysis, expression in antisense orientation in tomato plants
-
genotyping in diverse cancer cell lines, overview
-
His-tagged enzyme expression in Escherichia coli strain BL21-Gold(DE3)
-
overexpression of the wild-type and a mutant enzyme, the latter containing the wild-type 2 subunits alpha and beta plus an additional third one, gamma, overexpression of the 3-subunit mutant and the same mutant with point mutations of Arg-residues in Escherichia coli
-
wild-type and mutant enzymes are expressed in a idh1DELTAidh2DELTA yeast strain which contains deletion/insertion mutations in genomic IDH1 and IDH2 loci; wild-type and mutant enzymes are expressed in a idh1DELTAidh2DELTA yeast strain which contains deletion/insertion mutations in genomic IDH1 and IDH2 loci
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C201M/C332Y/K344D/Y345I/V351A/Y391K/R395S
-
converts the cofactor specificity from 7000-fold preference of NADP+ to a 200-fold preference of NAD+
alphaY126E
-
site-directed mutagenesis of subunit alpha, almost inactive mutant, shows low activity at pH 6.1 instead of pH 7.2, Km,Mn2+ is 30fold higher in the alphaY126E mutant as compared with the wild-type. Km,NAD+ for the alphaY126E mutant is 29fold higher than that of the wild-type. The Vmax of the wild-type at pH 6.1 is 0.0144 mmol/min/mg, whereas that for the alphaY126E mutant is only 0.00103 mmol/min/mg, suggesting a critical role for the residue in enzyme activity
alphaY126F
-
site-directed mutagenesis of subunit alpha, inactive mutant
alphaY126S
-
site-directed mutagenesis of subunit alpha, inactive mutant
betaY137E
-
site-directed mutagenesis of subunit beta, the mutant shows reduced activity compared to the wild-type enzyme
betaY137F
-
site-directed mutagenesis of subunit beta, the mutant shows reduced activity compared to the wild-type enzyme
betaY137S
-
site-directed mutagenesis of subunit beta, the mutant shows reduced activity compared to the wild-type enzyme
D181N
-
mutation in the alpha-subunit exhibits a 2000fold decrease in Vmax, with increases of 15fold in the Kms for Mn2+ and NAD+ and a much smaller change in the Km for isocitrate. Mutant enzyme fails to respond to ADP by lowering the Km for isocitrate, although it binds to ADP
D190N
-
mutation in the gamma-subunit results in 4-5fold decrease in Vmax as compared to wild-type enzyme. The Km-values for NAD+ and for Mn2+ of the mutant enzyme are 19 and 72 times, respectively, that of the wild-type enzyme with a much smaller effect on the Km for isocitrate. Mutant enzyme fails to respond to ADP by lowering the Km for isocitrate, although it binds to ADP
D192N
-
mutation in the beta-subunit results in 4-5fold decrease in Vmax as compared to wild-type enzyme. The Km-value for NAD+ of the mutant enzyme is 9times that of the normal enzyme with little or no effect on the affinity for Mn2+ or isocitrate. Mutant enzyme fails to respond to ADP by lowering the Km for isocitrate, although it binds to ADP
D206N
-
alpha-subunit mutant; specific activity of the alpha-subunit mutant enzyme is 1.3fold higher than specific activity of wild-type enzyme
D215N
-
gamma-subunit mutant; specific activity of the gamma-subunit mutant enzyme is 17% of specific activity of wild-type enzyme. 100fold increase in KM-value for Mn2+. Km-value for isocitrate is elevated
D217N
-
beta-subunit mutant; specific activity of the beta-subunit mutant enzyme is 33% of specific activity of wild-type enzyme. 16fold increase in KM-vlaue for NAD+
D230C
-
alpha-subunit mutant; specific activity of the alpha-subunit mutant enzyme is 6% of specific activity of wild-type enzyme. 32fold increase in KM-value for Mn2+. Km-value for isocitrate is elevated. 16fold increase in KM-vlaue for NAD+
D230N
-
alpha-subunit mutant; mutation in alpha-subunit results in complete loss of activity
D234C
-
alpha-subunit mutant; specific activity of the alpha-subunit mutant enzyme is 1% of specific activity of wild-type enzyme
D234N
-
alpha-subunit mutant; mutation in alpha-subunit results in complete loss of activity
gammaY135F
-
site-directed mutagenesis of subunit gamma, inactive mutant
L98P
the mutation is associated with retinitis pigmentosa
R132C
-
naturally occuring IDH1 mutation
R132G
-
naturally occuring IDH1 mutation
R132H
-
naturally occuring IDH1 mutation
R132L
-
naturally occuring IDH1 mutation
R132S
-
naturally occuring IDH1 mutation
R132V
-
naturally occuring IDH1 mutation
R88Q
-
site-directed mutagenesis, residue of the alpha-subunit, inactive mutant
R97Q
-
site-directed mutagenesis, residue of the additional mutant gamma-subunit, highly reduced activity compared to the wild-type enzyme
R99Q
-
site-directed mutagenesis, residue of the beta-subunit, reduced activity compared to the wild-type enzyme
D328K
-
the mutant shows dual coenzyme specificity
D328K/I329Y
-
introduction of the double mutation shifts the cofactor preference from NAD+ to NADP+
A108R/F136Y/T241D/N245D
-
site-directed mutagenesis, residues of subunit IDH1 mutant is unable to bind AMP, ligand-binding analysis
A108R/F136Y/T241D/N245D/R114A/Y142F/D248T/D252N
-
site-directed mutagenesis, residues of subunit IDH1 are A108R, F136Y, T241D, and N245D, residues of subunit IDH2 are R114A, Y142F, D248T, and D252N, mutant is unable to bind AMP, ligand-binding analysis
C56S/C150S/C242S
-
site-directed mutagenesis, IDH1/IDH2C150S octameric enzyme
C56S/C242S
D279A/D280A
-
site-directed mutagenesis, mutation of a IDH1 residue, reduced AMP binding, ligand-binding analysis
D286A/I287A
-
site-directed mutagenesis, mutation of a IDH2 residue, highly reduced NAD+ binding, ligand-binding analysis
H281A
-
site-directed mutagenesis, mutation of a IDH2 residue, mutant cannot bind NAD+, ligand-binding analysis
I221A
residue changes in IDH2 subunits
I221A/V225A/V229A
mutant enzyme IDH1-IDH2(I221A/V225A/V229A) shows an about 6fold decrease in apparent affinity for isocitrate measured in the absence or presence of AMP as compared to wild-type enzyme. Mutant enzyme IDH1-IDH2 (I221A/V225A/V229A) exhibits a moderate decrease in apparent affnity for cofactor (about 2.5fold relative to wild-type) in the absence of AMP. The mutant enzyme has acquired the novel property of cooperativity with respect to NAD+, but this cooperativity is not apparent in the presence of AMP. The mutant enzyme with residue changes in only one subunit are active in vivo; mutant enzyme IDH1-IDH2(I221A/V225A/V229A) shows an about 6fold decrease in apparent affinity for isocitrate measured in the absence or presence of AMP as compared to wild-type enzyme. Mutant enzyme IDH1-IDH2 (I221A/V225A/V229A) exhibits a moderate decrease in apparent affnity for cofactor (about 2.5fold relative to wild-type) in the absence of AMP. The mutant enzyme has acquired the novel property of cooperativity with respect to NAD+, but this cooperativity is not apparent in the presence of AMP. The mutant enzyme with residue changes in only one subunit are active in vivo
K171L
-
mutant IDH1K171L
R114A/Y142F/D248T/D252N
-
site-directed mutagenesis, residues of subunit subunit IDH2, ligand-binding analysis
R274A
-
site-directed mutagenesis, mutation of a IDH1 residue, reduced AMP binding, ligand-binding analysis
S220A
residue changes in IDH1 subunits
S92A/S98A
-
site-directed mutagenesis, residue of subunit IDH1 is S92, residue of subunit IDH2 is S98, ligand-binding analysis
V214A
residue changes in IDH1 subunits
V216A/S220A/V224A
IDH1(V216A/S220A/V224A)IDH2 mutant enzyme shows an about 6fold decrease in apparent affinity for isocitrate measured in the absence or presence of AMP as compared to wild-type enzyme. IDH1(V216A/S220A/V224A)IDH2 mutant enzyme exhibits a moderate decrease in apparent affnity for cofactor (about 2.5fold relative to wild-type) in the absence of AMP. The mutant enzyme has acquired the novel property of cooperativity with respect to NAD+, but this cooperativity is not apparent in the presence of AMP. The mutant enzyme with residue changes in only one subunit are active in vivo; IDH1(V216A/S220A/V224A)IDH2 mutant enzyme shows an about 6fold decrease in apparent affinity for isocitrate measured in the absence or presence of AMP as compared to wild-type enzyme. IDH1(V216A/S220A/V224A)IDH2 mutant enzyme exhibits a moderate decrease in apparent affnity for cofactor (about 2.5fold relative to wild-type) in the absence of AMP. The mutant enzyme has acquired the novel property of cooperativity with respect to NAD+, but this cooperativity is not apparent in the presence of AMP. The mutant enzyme with residue changes in only one subunit are active in vivo
V216A/S220A/V224A/I221A/V225A/V229A