Any feedback?
Please rate this page
(enzyme.php)
(0/150)

BRENDA support

BRENDA Home
show all | hide all No of entries

Information on EC 1.1.1.85 - 3-isopropylmalate dehydrogenase and Organism(s) Thermus thermophilus and UniProt Accession Q72IW9

for references in articles please use BRENDA:EC1.1.1.85
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
EC Tree
IUBMB Comments
The product decarboxylates spontaneously to yield 4-methyl-2-oxopentanoate.
Specify your search results
Select one or more organisms in this record: ?
This record set is specific for:
Thermus thermophilus
UNIPROT: Q72IW9
Show additional data
Do not include text mining results
Include (text mining) results
Include results (AMENDA + additional results, but less precise)
Word Map
The taxonomic range for the selected organisms is: Thermus thermophilus
The enzyme appears in selected viruses and cellular organisms
Synonyms
3-isopropylmalate dehydrogenase, ipmdh, beta-isopropylmalate dehydrogenase, isopropylmalate dehydrogenase, ipmdh2, ipmdh3, beta-ipm dehydrogenase, sbipmdh, ipmdh1, 3-ipm dehydrogenase, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2-hydroxy-4-methyl-3-carboxyvalerate:NAD+ oxidoreductase
-
-
-
-
2R,3S-isopropylmalate:NAD+ oxidoreductase (decaboxylating)
-
-
-
-
3-IPM dehydrogenase
-
-
-
-
3-IPM-DH
-
-
-
-
3-isopropylmalate dehydrogenase
beta-IPM dehydrogenase
-
-
-
-
beta-IPMDH
-
-
-
-
beta-isopropylmalate dehydrogenase
-
-
-
-
beta-isopropylmalic enzyme
-
-
-
-
dehydrogenase, 3-isopropylmalate
-
-
-
-
IPMDH
isopropylmalate dehydrogenase
NAD-dependent isopropylmalate dehydrogenase
-
threo-Ds-3-isopropylmalate dehydrogenase
-
-
-
-
Tt-beta-3-isopropylmalateDH
-
two-domain 3-isopropylmalate dehydrogenase
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
(2R,3S)-3-isopropylmalate + NAD+ = (2S)-2-isopropyl-3-oxosuccinate + NADH + H+
show the reaction diagram
the oxidative decarboxylation of 2-hydroxy acid substrates by the enzyme occurs via a random kinetic mechanism with the formation of two abortive complexes: enzyme-NADH-substrate and enzyme-NADH-inhibitor
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
redox reaction
-
-
-
-
oxidation
-
-
-
-
reduction
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
(2R,3S)-3-isopropylmalate:NAD+ oxidoreductase
The product decarboxylates spontaneously to yield 4-methyl-2-oxopentanoate.
CAS REGISTRY NUMBER
COMMENTARY hide
9030-97-1
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(2R,3S)-3-isopropylmalate + NAD+
(2S)-2-isopropyl-3-oxosuccinate + NADH + H+
show the reaction diagram
mutant enzymes with increased activity
-
-
?
(2R,3S)-3-isopropylmalate + NAD+
(2S)-2-isopropyl-3-oxosuccinate + NADH + H+
show the reaction diagram
(2R,3S)-3-isopropylmalate + NAD+
2-oxoisocaproate + NADH + H+ + CO2
show the reaction diagram
(2R,3S)-3-isopropylmalate + NAD+
4-methyl-2-oxopentanoate + CO2 + NADH + H+
show the reaction diagram
-
-
-
?
3-isopropylmalate + NAD+
2-ketoisocaproate + NADH + CO2
show the reaction diagram
-
-
-
?
DL-3-isopropylmalate + NAD+
DL-isopropyl-3-oxosuccinate + NADH + H+
show the reaction diagram
-
-
-
-
?
malate + NAD+
pyruvate + NADH + H+ + CO2
show the reaction diagram
-
-
-
-
?
malate + NADP+
pyruvate + NADPH + CO2
show the reaction diagram
-
-
-
-
?
tartrate + NAD+
?
show the reaction diagram
-
-
-
-
?
tartrate + NADP+
?
show the reaction diagram
-
-
-
-
?
threo-D-3-isopropylmalate + NAD+
?
show the reaction diagram
domain 1 binds the coenzyme NAD+, while the substrate 3-isopropylmalate binds mainly to domain 2
-
-
?
threo-DL-isopropylmalate + NAD+
2-isopropyl-3-oxosuccinate + NADH
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
(2R,3S)-3-isopropylmalate + NAD+
(2S)-2-isopropyl-3-oxosuccinate + NADH + H+
show the reaction diagram
(2R,3S)-3-isopropylmalate + NAD+
2-oxoisocaproate + NADH + H+ + CO2
show the reaction diagram
(2R,3S)-3-isopropylmalate + NAD+
4-methyl-2-oxopentanoate + CO2 + NADH + H+
show the reaction diagram
-
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Rb+
-
RbCl enhances activity
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(2S,3S)-3-methylmercaptomalate
strong competitive inhibitor
1,2-Cyclohexanediamine-N,N,N',N'-tetraacetate
-
-
isocitrate
-
-
NADH
-
NADH always produces competitive inhibition patterns with respect to NAD and noncompetitive inhibition patterns with respect to the substrates
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.59 - 2.6
(2R,3S)-3-isopropylmalate
0.9 - 1.9
NAD+
0.013 - 0.255
(2R,3S)-3-isopropylmalate
0.002 - 0.0133
DL-3-isopropylmalate
0.000017 - 0.316
isopropylmalate
0.0326 - 35.59
malate
0.0022 - 3.56
NAD+
0.014 - 6.579
NADP+
0.0407 - 1.21
Tartrate
0.0061 - 0.0161
threo-D,L-isopropylmalate
0.08
threo-Ds-3-Isopropylmalate
-
60°C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.08 - 5.7
(2R,3S)-3-isopropylmalate
0.0025 - 220
(2R,3S)-3-isopropylmalate
0.09 - 19
isopropylmalate
0.19 - 10.6
malate
0.15 - 220
NAD+
0.09 - 4.69
NADP+
0.1 - 0.26
Tartrate
additional information
additional information
-
the cold adaption results from the destabilization of the ternary complex caused by increase in the volume of the residue at position 126
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.16 - 5.4
(2R,3S)-3-isopropylmalate
0.12 - 4.2
NAD+
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.000062
(2S,3S)-3-methylmercaptomalate
pH 7.8, 60°C
additional information
additional information
-
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.2
-
at 75°C, in presence of K+
9
-
75°C, without K+
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
Swissprot
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
the enzyme is involved in the third step of the leucine biosynthesis pathway
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
73300
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
homodimer
additional information
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
crystal structure of mutant H15Y/E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M is determined at 2.4 A
ammonium sulfate precipitation, crystals diffract beyond 2.5 A resolution and are quite stable against X-rays, recombinant enzyme expressed in Escherichia coli
-
apo-form without substrate and in complex with the divalent metal–ion, in complexes with both Mn2+ and 3-isopropylmalate, as well as with both Mn2+ and NADH, at resolutions ranging from 1.8 to 2.5 A. Identification of two hinges at the interdomain region, hinge 1 between alphad and betaF as well as hinge 2 between alphah and betaE with a possible operational mechanism upon the action of the substrates. The interactions of the protein with Mn2+ and isopropylmalate are mainly responsible for the domain closure. Upon binding into the cleft of the interdomain region, the substrate isopropylmalate induces a relative movement of the secondary structural elements betaE, betaF, betaG, alphad and alphah. A movement of the loop bearing the amino acid Tyr139 precedes the interacting arm of the subunit. The tyrosyl ring rotates and moves by at least 5 A upon substrate binding. Thereby, new hydrophobic interactions are formed above the buried isopropyl-group of isopropylmalate. Domain closure is then completed only through subunit interactions. A loop of one subunit that is inserted into the interdomain cavity of the other subunit extends the area with the hydrophobic interactions, providing an example of the cooperativity between interdomain and intersubunit interactions
crystal structure of Thermus thermophilus IPMDH in a ternary complex with NAD+ and the inhibitor ((2S,3S)-3-methylmercaptomalate) is determined at 2.8 A resolution. The inhibitor exists as a decarboxylated product with an enol/enolate form in the active site. The product interacts with Arg94, Asn102, Ser259, Glu270, and a water molecule hydrogen-bonding with Arg132. All interactions between the product and the enzyme are observed in the position associated with keto-enol tautomerization
enzyme in complex with NAD+
-
hanging drop vapor diffusion method, chimeric enzyme constructed by fusing the gene of Bacillus subtilis and Thermus thermophilus coding for 3-isopropylmalate dehydrogenase, expression in Escherichia coli
-
in complex with Mn2+, (2R,3S)-3-isopropylmalate, and NADH, hanging drop vapor diffusion method, using 20% (w/v) PEG 6000, 0.1 M MOPS/KOH, pH 7.6, and 10% (v/v) ethanol
-
mutant enzyme E270A, hanging drop vapor diffusion method, using 20% (w/v) PEG 6000, 0.1 M MOPS/KOH pH 7.6, and 10% (v/v) ethanol
mutant enzymes A172V, A172G and A172F. As in the case of A172L enzyme, the A172F mutant can not be crystallized by the salting-out technique with ammonium sulfate. The crystal is obtained at pH 4.8 using polyethylene glycol 4000 as a precipitant. Crystals of mutant enzyme A172E are obtained from a drop equilibrated with reservoir solution consisting of 0.8 M ammonium sulfate pH 6.0 at either 15°C or 20°C K. Two types of crystals: one hexagonal bipyramidal and the other is tetragonal
-
recombinant enzyme expressed in Escherichia coli, ammonium sulfate precipitation
-
study of the mutant enzymes G240A and L246E/V249M by X-ray crystallography
-
tertiary structure solved by X-ray crystallography at 2.2 A resolution
using the hanging-drop vapour diffusion method crystals of Tt-IPMDH are grown in the following combinations: Apo Tt-IPMDH (crystal diffracted to a resolution of about 1.8 A), Tt-IPMDH in complex with Mn2+ (crystal diffracted to a resolution of about 2.5 A), Tt-IPMDH in complex with Mn2+ and beta-3-isopropylmalate (crystal diffracted to a resolution of 2.2 A), Tt-IPMDH in complex with Mn2+ and NADH (crystal diffracted to a resolution of 2.5 A), and Tt-IPMDH in complex with Mn2+, beta-3-isopropylmalate and NADH (crystal diffracted to a resolution of 2.75 A)
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E57V/R85M/Y86A/M208T/F217Y/V238M/R310M
no (2R,3S)-3-isopropylmalate dehydrogenase activity detected
E57V/S72I/R85M/Y86A/F217Y/V238M/R310M
no (2R,3S)-3-isopropylmalate dehydrogenase activity detected
E57V/S72I/R85M/Y86A/M208T/F217Y/R310M
in comparison to mutant H15Y/E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M this mutant shows a significant decrease in kcat ((2R,3S)-3-isopropylmalate), alteration of Km value only moderate
E57V/S72I/R85M/Y86A/M208T/F217Y/V238M
in comparison to mutant H15Y/E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M this mutant shows a significant decrease in kcat ((2R,3S)-3-isopropylmalate), alteration of Km value only moderate
E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M
in comparison to mutant H15Y/E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M this mutant shows a larger kcat ((2R,3S)-3-isopropylmalate) and a larger Km((2R,3S)-3-isopropylmalate) or (NAD+) temperatures giving half of the initial activity is 87.8 °C
E57V/S72I/R85M/Y86A/M208T/V238M/R310M
in comparison to mutant H15Y/E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M this mutant shows a significant decrease in kcat ((2R,3S)-3-isopropylmalate), alteration of Km value only moderate
E57V/S72I/R85V/Y86A/M208T/F217Y/V238M/R310M
no (2R,3S)-3-isopropylmalate dehydrogenase activity detected
H15Y/E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M
TtHICDH variant with considerably higher IPMDH activity than double mutant R85V/Y86A, Km ((2R,3S)-3-isopropylmalate) or (NAD+) lower than wild-type HICDH but still considerably higher than wild-type (2R,3S)-3-isopropylmalate dehydrogenase, kcat ((2R,3S)-3-isopropylmalate) much higher than wild-type HICDH but still considerably lower than wild-type (2R,3S)-3-isopropylmalate dehydrogenase, temperatures giving half of the initial activity is 79.9 °C, crystal structure determined at 2.4 A, mutant is a homotetramer
R85V/Y86A
TtHICDH variant with high IPMDH activity, Km ((2R,3S)-3-isopropylmalate) or (NAD+) lower than wild-type HICDH but still considerably higher than wild-type (2R,3S)-3-isopropylmalate dehydrogenase, kcat ((2R,3S)-3-isopropylmalate) higher than wild-type HICDH but still considerably lower than wild-type (2R,3S)-3-isopropylmalate dehydrogenase, temperatures giving half of the initial activity is 92.5 °C
S72I/R85M/Y86A/M208T/F217Y/V238M/R310M
in comparison to mutant H15Y/E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M this mutant shows a significant decrease in kcat ((2R,3S)-3-isopropylmalate), alteration of Km value only moderate
A172D
A172E
-
enhanced thermostability compared to wild-type enzyme
A172F
A172G
-
melting temperature is reduced by 0.3°C compared to wild-type enzyme, crystal structure is similar to that of the wild-type enzyme
A172I
-
melting temperature is increased by 2°C compared to wild-type enzyme
A172L
A172N
-
enhanced thermostability compared to wild-type enzyme
A172Q
-
enhanced thermostability compared to wild-type enzyme
A172V
A31G/G43A/A709G
-
optimum temperature is shifted to lower temperatures compared to wild-type enzyme, activity is less temperature dependent than that of the wild-type, resulting in higher activity at temperatures below 60°C, improved turnover-numbers at 40°Cm increase in Km-values, slight loss of thermal stability
A335E
C275T
-
similar pH-dependence to that of the wild-type enzyme, improved binding affinities for both D-3-isopropylmalate and NAD+ result in an improved catalytic efficiency, mutant enzyme retains thermal stability
D184H
D217A
the mutant retains 1.1% of the catalytic activity of the wild type enzyme
D241A
the mutant shows a drastic decrease in the kcat value (0.06% compared to that of the wild type enzyme)
D245A
the mutant retains 10.5% of the catalytic activity of the wild type enzyme
E270A
E87G
-
mutation does not affect the dissociation constant from the binary complex, but does greatly increase the KM-values
E87Q
-
mutation does not affect the dissociation constant from the binary complex, but does greatly increase the KM-values
G240A
-
decreased thermostability
G376A
-
optimum temperature is shifted to lower temperatures compared to wild-type enzyme, activity is less temperature dependent than that of the wild-type, resulting in higher activity at temperatures below 60°C, improved turnover-numbers at 40°C, increase in Km-values, mutant enzyme retains thermal stability
G43A
-
optimum temperature is shifted to lower temperatures compared to wild-type enzyme, activity is less temperature dependent than that of the wild-type, resulting in higher activity at temperatures below 60°C, improved turnover-numbers at 40°C, increase ion Km-values, slight loss of thermal stability
H179K
K185A
the mutant shows a drastic decrease in the kcat value (0.06% compared to that of the wild type enzyme)
L134N
L134N/V181T/P324T/A335E
site-directed mutagenesis, exchange of residues for those of ancestral mutants, the mutant shows increased thermal stability compared to the wild-type enzyme, the mutant shows increased thermal stability compared to the wild-type enzyme
L204F
-
higher thermostability compared to wild-type enzyme, denaturation rates at 68°C and 70°C are slower
L246E/V249M
-
decreased thermostability
N102A
the mutant retains 13% of the catalytic activity of the wild type enzyme
P324T
S226R
-
Km-value for NADP+ is about 40% compared to wild-type enzyme, Km-value for NAD+ is 2.6fold higher compared to wild-type enzyme
S226R/D278K/I279Y
-
Km-value for NADP+ is about 0.8% compared to wild-type enzyme, Km-value for NAD+ is 153fold higher compared to wild-type enzyme
S261N
T16V
site-directed mutagenesis, exchange of a cofactor binding residue to the residue of the ancestral mutant, the mutant shows increased thermal stability compared to the wild-type enzyme
V126A
-
the mutation decreases the KM value of DL-3-isopropylmalate at 60°C in comparison to wild-type
V126E
-
the mutation increases the KM value of DL-3-isopropylmalate at 60°C in comparison to wild-type
V126F
-
the mutation increases the KM value of DL-3-isopropylmalate at 60°C in comparison to wild-type
V126G
-
the mutation decreases the KM value of DL-3-isopropylmalate at 60°C in comparison to wild-type
V126I
-
the mutation increases the KM value of DL-3-isopropylmalate at 60°C in comparison to wild-type
V126L
-
the mutation increases the KM value of DL-3-isopropylmalate at 60°C in comparison to wild-type
V126M
-
the mutation increases the KM value of DL-3-isopropylmalate at 60°C in comparison to wild-type
V126S
-
the mutation decreases the KM value of DL-3-isopropylmalate at 60°C in comparison to wild-type
V126T
-
the mutation decreases the KM value of DL-3-isopropylmalate at 60°C in comparison to wild-type
V181T
V181T/P324T/A335E
site-directed mutagenesis, exchange of residues for those of ancestral mutants, the mutant highly shows increased thermal stability compared to the wild-type enzyme, the mutant shows decreased thermal stability compared to the wild-type enzyme
W152F
site-directed mutagenesis, the W152F mutation causes a significant decrease in the amplitude of only the slow part of refolding, without influencing the amplitude of the burst part, the mutant shows an altered tertiary structure compared to the wild-type enzyme conformation
W152F/W195F
site-directed mutagenesis, the W152F mutation causes a significant decrease in the amplitude of only the slow part of refolding, without influencing the amplitude of the burst part, the mutant shows an altered tertiary structure compared to the wild-type enzyme conformation
W77F/W152F
site-directed mutagenesis, the W152F mutation causes a significant decrease in the amplitude of only the slow part of refolding, without influencing the amplitude of the burst part, the mutant shows an altered tertiary structure compared to the wild-type enzyme conformation
Y139A
the mutant retains 2.9% of the catalytic activity of the wild type enzyme
additional information
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50
thermal stability curves of wild-type and mutant enzymes at pH 7.6 in presence of 0.5 mM EDTA, overview
61
-
midpoint of thermal unfolding curve, mutant enzyme A172D
65
-
midpoint of thermal unfolding curve, mutant enzyme A172V
66
-
midpoint of thermal unfolding curve, mutant enzyme A172I and A172E
67
-
midpoint of thermal unfolding curve, mutant enzyme A172L and A172F
79
-
midpoint of irreversible denaturation
82
-
10 min, 50% inactivation, mutant enzyme A31G/G43A/A709G
83
-
10 min, 50% inactivation
85
-
10 min, 50% inactivation, mutant enzyme G43A
85 - 95
thermal degeneration curves of wild-type and mutant enzymes at pH 7.6, overview
additional information
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
complete loss of activity in presence of 4 M urea
-
higher thermal stability of the dimer as compared to monomer. B24-B24' is the major contributor to maintaining subunit-subunit interaction at 64°C
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, stable for at least 6 months when frozen quickly, 40% inactivation when frozen slowly
-
4°C , 50 mM potassium phosphate buffer pH 7.6, 0.5 mM EDTA, 6 months, stable
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene cloned in Escherichia coli
-
recombinant wild-type and mutants from Escherichia coli strain MA153 by anion exchange and adsorption chromatography
using Ni-NTA chromatography
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli
construction of a chimeric gene by fusing the Bacillus subtilis and Thermus thermophilus genes coding for 3-isopropylmalate dehydrogenase, expression in Escherichia coli
-
expressed as His-tagged and non-His-tagged fusion proteins in Escherichia coli
expressed in Escherichia coli as a His-tagged fusion protein
expression in Escherichia coli
-
expression in Escherichia coli using pBR322 as a vector
-
expression of wild-type and mutant enzymes in Escherichia coli
-
gene leuB, DNA and amino acid sequence determination and analysis, phylogenetic analysis, expression of wild-type and mutants in Escherichia coli strain MA153
gene leuB, expression of wild-type and mutants in Escherichia coli strain MA153
phylogenetic analysis, comparison to isocitrate dehydrogenase, EC 1.1.1.41, overview
the sequence motif is introduced into a mesophilic Escherichia coli isopropylmalate dehydrogenase, one by one. The introduction of the whole motif leads the mesophilic enzyme to be more unstable whereas substitution of only one amino acid residue in the motif thermostabilizes the enzyme
-
RENATURED/Commentary
ORGANISM
UNIPROT
LITERATURE
enzyme solution containing 3.2 M urea is diluted 10times with a urea-free buffer. In samples containing 0.2 mM MnCl2 the activity is restored to 55-60%, full activity is recovered in absence of MnCl2
-
refolding mechanism of the homodimeric enzyme, dilution of the denatured protein, overview. Association of the two polypeptide chains occurs at the beginning of refolding, overview
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Onodera, K.; Moriyama, H.; Takenaka, A.; Tanaka, N.; Akutsu, N.; Muro, M.; Oshima, T.; Imada, K.; Sato, M.; Katsube, Y.
Crystallization and preliminary X-ray studies of a Bacillus subtilis and Thermus thermophilus HB8 chimeric 3-isopropylmalate dehydrogenase
J. Biochem.
109
1-2
1991
Bacillus subtilis, Thermus thermophilus
Manually annotated by BRENDA team
Yamada, T.; Akutsu, N.; Miyazaki, K.; Kakinuma, K.; Yoshida, M.; Oshima, T.
Purification, catalytic properties, and thermal stability of threo-Ds-3-isopropylmalate dehydrogenase coded by leuB gene from an extreme thermophile, Thermus thermophilus strain HB8
J. Biochem.
108
449-456
1990
Thermus thermophilus
Manually annotated by BRENDA team
Kakinuma, K.; Ozawa, K.; Fujimoto, Y.; Akutsu, N.; Oshima, T.
Stereochemistry of the decarboxylation reaction catalysed by 3-isopropylmalate dehydrogenase from the thermophilic bacterium Thermus thermophilus
J. Chem. Soc. Chem. Commun.
1989
1190-1192
1989
Thermus thermophilus
-
Manually annotated by BRENDA team
Katsube, Y.; Tanaka, N.; Takenaka, A.; Yamada, T.; Oshima, T.
Crystallization and preliminary X-ray data for 3-isopropylmalate dehydrogenase of Thermus thermophilus
J. Biochem.
104
679-680
1988
Thermus thermophilus
Manually annotated by BRENDA team
Hamasawa, K.; Kobayashi, Y.; Harada, S.; Yoda, K.; Yamasaki, M.; Tamura, G.
Molecular cloning and nucleotide sequence of the 3-isopropylmalate dehydrogenase gene of Candida utilis
J. Gen. Microbiol.
133
1089-1097
1987
Cyberlindnera jadinii, Saccharomyces cerevisiae, Thermus thermophilus
Manually annotated by BRENDA team
Sekiguchi, T.; Suda, M.; Sekiguchi, T.; Nosoh, Y.
Cloning and DNA homology of 3-isopropylmalate dehydrogenase genes from thermophilic bacilli
FEMS Microbiol. Lett.
36
41-45
1986
[Bacillus] caldolyticus, [Bacillus] caldotenax, Bacillus subtilis, Saccharomyces cerevisiae, Escherichia coli, Thermus thermophilus
-
Manually annotated by BRENDA team
Sekiguchi, T.; Ortega-Cesena, J.; Nosoh, Y.; Ohashi, S.; Tsuda, K.; Kanaya, S.
DNA and amino-acid sequences of 3-isopropylmalate dehydrogenase of Bacillus coagulans. Comparison with the enzymes of Saccharomyces cerevisiae and Thermus thermophilus
Biochim. Biophys. Acta
867
36-44
1986
Weizmannia coagulans, Bacillus subtilis, Saccharomyces cerevisiae, Thermus thermophilus
-
Manually annotated by BRENDA team
Tanaka, T.; Kawano, N.; Oshima, T.
Cloning of 3-isopropylmalate dehydrogenase gene of an extreme thermophile and partial purification of the gene product
Biochemistry
89
677-682
1981
Thermus thermophilus
Manually annotated by BRENDA team
Croft, J.E.; Love, D.R.; Bergquist, P.L.
Expression of leucine genes from an extremely thermophilic bacterium in Escherichia coli
Mol. Gen. Genet.
210
490-497
1987
Escherichia coli, Thermus thermophilus, Salmonella enterica subsp. enterica serovar Typhimurium
Manually annotated by BRENDA team
Wallon, G.; Kryger, G.; Lovett, S.T.; Oshima, T.; Ringe, D.; Petsko, G.A.
Crystal structures of Escherichia coli and Salmonella typhimurium 3-isopropylmalate dehydrogenase and comparison with their thermophilic counterpart from Thermus thermophilus
J. Mol. Biol.
266
1016-1031
1997
Escherichia coli, Salmonella enterica subsp. enterica serovar Typhimurium, Thermus thermophilus
Manually annotated by BRENDA team
Dean, A.M.; Dvorak, L.
The role of glutamate 87 in the kinetic mechanism of Thermus thermophilus isopropylmalate dehydrogenase
Protein Sci.
4
2156-2167
1995
Thermus thermophilus
Manually annotated by BRENDA team
Suzuki, T.; Yasugi, M.; Arisaka, F.; Yamagishi, A.; Oshima, T.
Adaptation of a thermophilic enzyme, 3-isopropylmalate dehydrogenase, to low temperatures
Protein Eng.
14
85-91
2001
Thermus thermophilus
Manually annotated by BRENDA team
Aoshima, M.; Oshima, T.
Stabilization of Escherichia coli isopropylmalate dehydrogenase by single amino acid substitution
Protein Eng.
10
249-254
1997
Escherichia coli, Thermus thermophilus
Manually annotated by BRENDA team
Akanuma, S.; Qu, C.; Yamagishi, A.; Tanaka, N.; Oshima, T.
Effect of polar side chains at position 172 on thermal stability of 3-isopropylmalate dehydrogenase from Thermus thermophilus
FEBS Lett.
410
141-144
1997
Thermus thermophilus
Manually annotated by BRENDA team
Chen, R.; Greer, A.; Dean, A.M.
Redesigning secondary structure to invert coenzyme specificity in isopropylmalate dehydrogenase
Proc. Natl. Acad. Sci. USA
93
12171-12176
1996
Thermus thermophilus
Manually annotated by BRENDA team
Qu, C.; Akanuma, S.; Tanaka, N.; Moriyama, H.; Oshima, T.
Design, X-ray crystallography, molecular modelling and thermal stability studies of mutant enzymes at site 172 of 3-isopropylmalate dehydrogenase from Thermus thermophilus
Acta Crystallogr. Sect. D
57
225-232
2001
Thermus thermophilus
Manually annotated by BRENDA team
Wallon, G.; Yamamoto, K.; Kirino, H.; Yamagishi, A.; Lovett, S.T.; Petsko, G.A.; Oshima, T.
Purification, catalytic properties and thermostability of 3-isopropylmalate dehydrogenase from Escherichia coli
Biochim. Biophys. Acta
1337
105-112
1997
Escherichia coli, Thermus thermophilus
Manually annotated by BRENDA team
Hurley, J.H.; Dean, A.M.
Structure of 3-isopropylmalate dehydrogenase in complex with NAD+: ligand-induced loop closing and mechanism for cofactor specificity
Structure
2
1007-1016
1994
Thermus thermophilus
Manually annotated by BRENDA team
Moriyma, H.; Onodera, K.; Sakurai, M.; Tanaka, N.; Kirino-Kagawa, H.; Oshima, T.; Katsube, Y.
The crystal structures of mutated 3-isopropylmalate dehydrogenase from Thermus thermophilus HB8 and their relationship to the thermostability of the enzyme
J. Biochem.
117(2)
408-413
1995
Thermus thermophilus
-
Manually annotated by BRENDA team
Suzuki, T.; Yasugi, M.; Arisaka, F.; Oshima, T.; Yamagishi, A.
Cold-adaptation mechanism of mutant enzymes of 3-isopropylmalate dehydrogenase from Thermus thermophilus
Protein Eng.
15
471-476
2002
Thermus thermophilus
Manually annotated by BRENDA team
Watanabe, K.; Yamagishi, A.
The effects of multiple ancestral residues on the Thermus thermophilus 3-isopropylmalate dehydrogenase
FEBS Lett.
580
3867-3871
2006
Thermus thermophilus (Q5SIY4), Thermus thermophilus
Manually annotated by BRENDA team
Watanabe, K.; Ohkuri, T.; Yokobori, S.; Yamagishi, A.
Designing thermostable proteins: ancestral mutants of 3-isopropylmalate dehydrogenase designed by using a phylogenetic tree
J. Mol. Biol.
355
664-674
2006
Thermus thermophilus (Q5SIY4), Thermus thermophilus
Manually annotated by BRENDA team
Kalinina, O.V.; Gelfand, M.S.
Amino acid residues that determine functional specificity of NADP- and NAD-dependent isocitrate and isopropylmalate dehydrogenases
Proteins
64
1001-1009
2006
Acidithiobacillus ferrooxidans (Q56268), Agrobacterium tumefaciens (P24404), Arthrospira platensis (Q00412), Aspergillus niger (P87256), Azotobacter vinelandii (P96197), Bacteroides fragilis (P54354), Brassica napus (P29102), Candida maltosa (P07139), Clostridium pasteurianum (P31958), Cyberlindnera jadinii (P08791), Eremothecium gossypii (O60027), Escherichia coli (P30125), Leptospira interrogans (P24015), Mycobacterium tuberculosis variant bovis (P94929), Ogataea angusta (P34733), Saccharomyces cerevisiae (P87267), Salmonella enterica subsp. enterica serovar Typhimurium (P37412), Scheffersomyces stipitis (O94114), Thermus aquaticus (P24098), Thermus thermophilus (P61494), Thermus thermophilus (P61495), Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039 (P61494), Zymoseptoria tritici (Q9Y897), [Candida] boidinii (Q01987)
Manually annotated by BRENDA team
Graczer, E.; Varga, A.; Hajdu, I.; Melnik, B.; Szilagyi, A.; Semisotnov, G.; Zavodszky, P.; Vas, M.
Rates of unfolding, rather than refolding, determine thermal stabilities of thermophilic, mesophilic, and psychrotrophic 3-isopropylmalate dehydrogenases
Biochemistry
46
11536-11549
2007
Thermus thermophilus (Q5SIY4)
Manually annotated by BRENDA team
Graczer, E.; Varga, A.; Melnik, B.; Semisotnov, G.; Zavodszky, P.; Vas, M.
Symmetrical refolding of protein domains and subunits: example of the dimeric two-domain 3-isopropylmalate dehydrogenases
Biochemistry
48
1123-1134
2009
Vibrio sp., Escherichia coli (P30125), Escherichia coli, Thermus thermophilus (Q5SIY4), Thermus thermophilus, Vibrio sp. I5
Manually annotated by BRENDA team
Merli, A.; Manikandan, K.; Graczer, E.; Schuldt, L.; Singh, R.K.; Zavodszky, P.; Vas, M.; Weiss, M.S.
Crystallization and preliminary X-ray diffraction analysis of various enzyme-substrate complexes of isopropylmalate dehydrogenase from Thermus thermophilus
Acta Crystallogr. Sect. F
66
738-743
2010
Thermus thermophilus (Q5SIY4), Thermus thermophilus
Manually annotated by BRENDA team
Suzuki, Y.; Asada, K.; Miyazaki, J.; Tomita, T.; Kuzuyama, T.; Nishiyama, M.
Enhancement of latent 3-isopropylmalate dehydrogenase activity of promiscuous homoisocitrate dehydrogenase by directed evolution
Biochem. J.
431
401-410
2010
Thermus thermophilus (Q72IW9)
Manually annotated by BRENDA team
Nango, E.; Yamamoto, T.; Kumasaka, T.; Eguchi, T.
Crystal structure of 3-isopropylmalate dehydrogenase in complex with NAD(+) and a designed inhibitor
Bioorg. Med. Chem.
17
7789-7794
2009
Thermus thermophilus (Q5SIY4), Thermus thermophilus
Manually annotated by BRENDA team
Graczer, E.; Lionne, C.; Zavodszky, P.; Chaloin, L.; Vas, M.
Transient kinetic studies reveal isomerization steps along the kinetic pathway of Thermus thermophilus 3-isopropylmalate dehydrogenase
FEBS J.
280
1764-1772
2013
Thermus thermophilus
Manually annotated by BRENDA team
Graczer, E.; Konarev, P.V.; Szimler, T.; Bacso, A.; Bodonyi, A.; Svergun, D.I.; Zavodszky, P.; Vas, M.
Essential role of the metal-ion in the IPM-assisted domain closure of 3-isopropylmalate dehydrogenase
FEBS Lett.
585
3297-3302
2011
Thermus thermophilus
Manually annotated by BRENDA team
Graczer, E.; Merli, A.; Singh, R.K.; Karuppasamy, M.; Zavodszky, P.; Weiss, M.S.; Vas, M.
Atomic level description of the domain closure in a dimeric enzyme: Thermus thermophilus 3-isopropylmalate dehydrogenase
Mol. Biosyst.
7
1646-1659
2011
Thermus thermophilus (Q5SIY4), Thermus thermophilus
Manually annotated by BRENDA team
Graczer, E.; Szimler, T.; Garamszegi, A.; Konarev, P.V.; Labas, A.; Olah, J.; Pallo, A.; Svergun, D.I.; Merli, A.; Zavodszky, P.; Weiss, M.S.; Vas, M.
Dual role of the active site residues of Thermus thermophilus 3-isopropylmalate dehydrogenase: chemical catalysis and domain closure
Biochemistry
55
560-574
2016
Thermus thermophilus (Q5SIY4), Thermus thermophilus
Manually annotated by BRENDA team
Pallo, A.; Olah, J.; Graczer, E.; Merli, A.; Zavodszky, P.; Weiss, M.S.; Vas, M.
Structural and energetic basis of isopropylmalate dehydrogenase enzyme catalysis
FEBS J.
281
5063-5076
2014
Thermus thermophilus
Manually annotated by BRENDA team
Graczer, E.; Pallo, A.; Olah, J.; Szimler, T.; Konarev, P.V.; Svergun, D.I.; Merli, A.; Zavodszky, P.; Weiss, M.S.; Vas, M.
Glutamate 270 plays an essential role in K+-activation and domain closure of Thermus thermophilus isopropylmalate dehydrogenase
FEBS Lett.
589
240-245
2015
Thermus thermophilus (Q5SIY4), Thermus thermophilus, Thermus thermophilus HB8 / ATCC 27634 / DSM 579 (Q5SIY4)
Manually annotated by BRENDA team
Sharma, R.; Sastry, G.N.
Deciphering the dynamics of non-covalent interactions affecting thermal stability of a protein: molecular dynamics study on point mutant of Thermus thermophilus isopropylmalate dehydrogenase
PLoS ONE
10
e0144294
2015
Thermus thermophilus (Q5SIY4), Thermus thermophilus, Thermus thermophilus HB8 / ATCC 27634 / DSM 579 (Q5SIY4)
Manually annotated by BRENDA team
Kotsuka, T.; Akanuma, S.; Tomuro, M.; Yamagishi, A.; Oshima, T.
Further stabilization of 3-isopropylmalate dehydrogenase of an extreme thermophile, Thermus thermophilus, by a suppressor mutation method
J. Bacteriol.
178
723-727
1996
Thermus thermophilus (P61494), Thermus thermophilus
Manually annotated by BRENDA team
Sharma, R.; Sagurthi, S.; Narahari Sastry, G.
Elucidating the preference of dimeric over monomeric form for thermal stability of Thermus thermophilus isopropylmalate dehydrogenase A molecular dynamics perspective
J. Mol. Graph. Model.
96
107530
2020
Thermus thermophilus (Q5SIY4), Thermus thermophilus, Thermus thermophilus ATCC 27634 (Q5SIY4)
Manually annotated by BRENDA team
Akanuma, S.; Bessho, M.; Kimura, H.; Furukawa, R.; Yokobori, S.; Yamagishi, A.
Establishment of mesophilic-like catalytic properties in a thermophilic enzyme without affecting its thermal stability
Sci. Rep.
9
9346
2019
Thermus thermophilus (Q5SIY4), Thermus thermophilus
Manually annotated by BRENDA team