1.1.1.41: isocitrate dehydrogenase (NAD+)

This is an abbreviated version, for detailed information about isocitrate dehydrogenase (NAD+), go to the full flat file.

Reaction

isocitrate
+
NAD+
=
2-oxoglutarate
+
CO2
+
NADH

Synonyms

At2g17130, At4g35260, At4g35650, At5g03290, AtIDH I, AtIDH II, AtIDH V, AtIDH VI, beta-decarboxylating dehydrogenase, beta-ketoglutaric-isocitric carboxylase, Icd-2, ICDH, ICDH1, IDH, IDH I, IDH II, IDH III, IDH IV, IDH V, IDH-I, IDH-II, IDH-V, IDH1, IDH2, IDH5, IDHa, isocitrate dehydrogenase, isocitrate dehydrogenase 2, isocitrate-homoisocitrate dehydrogenase, isocitric acid dehydrogenase, isocitric dehydrogenase, McIDH, mIDH, mitochondrial NAD-dependent isocitrate dehydrogenase, NAD dependent isocitrate dehydrogenase, NAD isocitrate dehydrogenase, NAD isocitric dehydrogenase, NAD(P)+-dependent isocitrate dehydrogenase, NAD+-dependent ICDH, NAD+-dependent isocitrate dehydrogenase, NAD+-dependent isocitrate dehydrogenase 1, NAD+-specific ICDH, NAD+-specific IDH, NAD+-specific isocitrate dehydrogenase, NAD-Dependent IDH, NAD-dependent isocitrate dehydrogenase, NAD-ICDH, NAD-IDH, NAD-isocitrate dehydrogenase, NAD-linked isocitrate dehydrogenase, NAD-specific isocitrate dehydrogenase, PF0202, PH1722, Rv0066c, threo-DS-isocitrate:NAD+ oxidoreductase (decarboxylating)

ECTree

     1 Oxidoreductases
         1.1 Acting on the CH-OH group of donors
             1.1.1 With NAD+ or NADP+ as acceptor
                1.1.1.41 isocitrate dehydrogenase (NAD+)

Engineering

Engineering on EC 1.1.1.41 - isocitrate dehydrogenase (NAD+)

Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
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
in the absence of AMP, the IDH1(V216A/S220A/V224A)-IDH2(I221A/V225A/V229A) mutant enzyme demonstrates an about 30fold decrease in apparent Vmax relative to wild-type and a loss of cooperativity with respect to isocitrate and, in the presence of AMP, the apparent affinity of the enzyme for isocitrate is 35fold lower than that of the wild-type enzyme. This mutant enzyme also exhibits a significant decrease in apparent affinity for NAD+ (about 30fold in the absence of AMP and about 10fold in the presence of AMP relative to wild-type). 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 demonstrates decreases in apparent affnity for isocitrate of about 12fold in the absence of AMP and of about 26fold in the presence of AMP relative to wild-type. For this mutant enzyme, with respect to isocitrate, allosteric activation by AMP and cooperativity in the absence of AMP are no longer apparen. The mutant enzyme is not fully functional in vivo; in the absence of AMP, the IDH1(V216A/S220A/V224A)-IDH2(I221A/V225A/V229A) mutant enzyme demonstrates an about 30fold decrease in apparent Vmax relative to wild-type and a loss of cooperativity with respect to isocitrate and, in the presence of AMP, the apparent affinity of the enzyme for isocitrate is 35fold lower than that of the wild-type enzyme. This mutant enzyme also exhibits a significant decrease in apparent affinity for NAD+ (about 30fold in the absence of AMP and about 10fold in the presence of AMP relative to wild-type). 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 demonstrates decreases in apparent affnity for isocitrate of about 12fold in the absence of AMP and of about 26fold in the presence of AMP relative to wild-type. For this mutant enzyme, with respect to isocitrate, allosteric activation by AMP and cooperativity in the absence of AMP are no longer apparen. The mutant enzyme is not fully functional in vivo
V224A
residue changes in IDH1 subunits
V225A
residue changes in IDH2 subunits
V229A
residue changes in IDH2 subunits
C150S
-
insensitive to treatment with diamide, the presence of the IDH2 Cys-150 residue (and perhaps the potential for disulfide bond formation) may contribute to cooperativity and may also limit maximal IDH activity
-
C56S/C242S
-
shows activity similar to wild type enzyme and is sensitive to treatment with diamide
-
D279A
-
IDH1D279A
-
D286A
-
IDH2D286A
-
I221A
-
residue changes in IDH2 subunits
-
I280A
-
IDH1I280A
-
K171L
-
mutant IDH1K171L
-
S220A
-
residue changes in IDH1 subunits
-
V214A
-
residue changes in IDH1 subunits
-
V224A
-
residue changes in IDH1 subunits
-
V225A
-
residue changes in IDH2 subunits
-
S102A
-
site-directed mutagenesis, the mutant shows decreased affinity for isocitrate and 3.3% of wild-type activity
S102G
-
site-directed mutagenesis, the mutant shows decreased affinity for isocitrate and 2.8% of wild-type activity
S102T
-
site-directed mutagenesis, the mutant shows decreased affinity for isocitrate and 16% of wild-type activity
S102Y
-
site-directed mutagenesis, the mutant shows decreased affinity for isocitrate and 1.1% of wild-type activity
additional information