Information on EC 1.2.4.4 - 3-methyl-2-oxobutanoate dehydrogenase (2-methylpropanoyl-transferring)

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

EC NUMBER
COMMENTARY hide
1.2.4.4
-
RECOMMENDED NAME
GeneOntology No.
3-methyl-2-oxobutanoate dehydrogenase (2-methylpropanoyl-transferring)
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
3-methyl-2-oxobutanoate + [dihydrolipoyllysine-residue (2-methylpropanoyl)transferase] lipoyllysine = [dihydrolipoyllysine-residue (2-methylpropanoyl)transferase] S-(2-methylpropanoyl)dihydrolipoyllysine + CO2
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
-
-
-
-
oxidative decarboxylation
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
(S)-3-methyl-2-oxopentanoate dehydrogenase (acylating)
-
-
2-oxoisovalerate decarboxylation to isobutanoyl-CoA
-
-
4-methyl-2-oxopentanoate dehydrogenase (acylating)
-
-
Biosynthesis of antibiotics
-
-
Biosynthesis of secondary metabolites
-
-
isoleucine metabolism
-
-
Metabolic pathways
-
-
NIL
-
-
pantothenate biosynthesis
-
-
Propanoate metabolism
-
-
Valine, leucine and isoleucine degradation
-
-
SYSTEMATIC NAME
IUBMB Comments
3-methyl-2-oxobutanoate:[dihydrolipoyllysine-residue (2-methylpropanoyl)transferase]-lipoyllysine 2-oxidoreductase (decarboxylating, acceptor-2-methylpropanoylating)
Contains thiamine diphosphate. It acts not only on 3-methyl-2-oxobutanaoate, but also on 4-methyl-2-oxopentanoate and (S)-3-methyl-2-oxopentanoate, so that it acts on the 2-oxo acids that derive from the action of transaminases on valine, leucine and isoleucine. It is a component of the multienzyme 3-methyl-2-oxobutanoate dehydrogenase complex in which multiple copies of it are bound to a core of molecules of EC 2.3.1.168, dihydrolipoyllysine-residue (2-methylpropanoyl)transferase, which also binds multiple copies of EC 1.8.1.4, dihydrolipoyl dehydrogenase. It does not act on free lipoamide or lipoyllysine, but only on the lipoyllysine residue in EC 2.3.1.168.
CAS REGISTRY NUMBER
COMMENTARY hide
9031-05-4
-
9082-72-8
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain DSM 3757
-
-
Manually annotated by BRENDA team
E2 component; rainbow trout, the enzyme is the E2 dihydrolipoyl transacylase subunit of the branched-chain alpha-keto acid dehydrogenase multienzyme complex BCKADH
SwissProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2-oxo-4-methylthiobutanoate + NAD+ + CoA
3-methylthiopropionyl-CoA + NADH + CO2
show the reaction diagram
2-oxo-beta-methylvalerate + [dihydrolipoyllysine-residue(2-methylpropanoyl)transferase]-lipoyllysine
?
show the reaction diagram
2-oxo-butyrate + [dihydrolipoyllysine-residue(2-methylpropanoyl)transferase]-lipoyllysine
?
show the reaction diagram
2-oxo-gamma-methylthiobutyrate + [dihydrolipoyllysine-residue(2-methylpropanoyl)transferase]-lipoyllysine
?
show the reaction diagram
2-oxo-glutarate + [dihydrolipoyllysine-residue(2-methylpropanoyl)transferase]-lipoyllysine
?
show the reaction diagram
2-oxo-isocaproate + NAD+ + CoA
3-methylbutanoyl-CoA + CO2 + NADH + H+
show the reaction diagram
2-oxo-isocaproate + [dihydrolipoyllysine-residue(2-methylpropanoyl)transferase]-lipoyllysine
?
show the reaction diagram
2-oxobutanoate + NAD+ + CoA
propionyl-CoA + CO2 + NADH
show the reaction diagram
2-oxoglutarate + NAD+ + CoA
3-carboxypropionyl-CoA + NADH + CO2
show the reaction diagram
2-oxoisovalerate + 2,6-dichlorophenol indophenol
? + CO2
show the reaction diagram
-
-
-
-
?
2-oxoisovalerate + 2,6-dichlorophenolindophenol + CoA
? + CO2 + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
-
-
-
-
?
2-oxoisovalerate + NAD+
? + NADH
show the reaction diagram
-
-
-
-
?
2-oxoisovalerate + [dihydrolipoyllysine-residue(2-methylpropanoyl)transferase] lipoyllysine
?
show the reaction diagram
-
-
-
-
?
2-oxoisovalerate + [dihydrolipoyllysine-residue(2-methylpropanoyl)transferase]-lipoyllysine
?
show the reaction diagram
2-oxoisovalerate + [dihydrolipoyllysine-residue(2-methylpropanoyl)transferase]-lipoyllysine
? + CO2
show the reaction diagram
-
-
-
-
?
2-oxoisovalerate + [dihydrolipoyllysine-residue(2-methylpropanoyl)transferase]-lipoyllysine
[dihydrolipoyllysine-residue (2-methylpropanoyl)transferase] S-butanoyl-dihydrolipoyllysine + CO2
show the reaction diagram
-
overall reaction of the branched-chain alpha-keto acid dehydrogenase multienzyme complex BCKADH
-
-
?
2-oxopentanoate + NAD+ + CoA
butanoyl-CoA + CO2 + NADH
show the reaction diagram
-
-
-
-
?
3-methyl-2-oxobutanoate + NAD+ + CoA
2-methylpropanoyl-CoA + CO2 + NADH
show the reaction diagram
3-methyl-2-oxobutanoate + NAD+ + CoA
2-methylpropanoyl-CoA + CO2 + NADH + H+
show the reaction diagram
-
-
-
-
?
3-methyl-2-oxobutanoate + [dihydrolipoyllysine-residue (2-methylpropanoyl)transferase] lipoyllysine
[dihydrolipoyllysine-residue (2-methylpropanoyl)transferase] S-(2-methylpropanoyl)dihydrolipoyllysine + CO2
show the reaction diagram
-
-
-
-
?
3-methyl-2-oxobutanoate + [dihydrolipoyllysine-residue(2-methylpropanoyl)transferase]-lipoyllysine
[dihydrolipoyllysine-residue (2-methylpropanoyl)transferase] S-(2-methylpropanoyl)dihydrolipoyllysine + CO2
show the reaction diagram
3-methyl-2-oxopentanoate + NAD+ + CoA
2-methylbutanoyl-CoA + CO2 + NADH
show the reaction diagram
3-methyl-2-oxopentanoate + [dihydrolipoyllysine-residue(2-methylpropanoyl)transferase]-lipoyllysine
[dihydrolipoyllysine-residue (2-methylpropanoyl)transferase] S-(2-methylbutanoyl)dihydrolipoyllysine + CO2
show the reaction diagram
-
-
-
-
r
4-methyl-2-oxopentanoate + NAD+ + CoA
3-methylbutanoyl-CoA + CO2 + NADH
show the reaction diagram
4-methyl-2-oxopentanoate + [dihydrolipoyllysine-residue(2-methylpropanoyl)transferase]-lipoyllysine
[dihydrolipoyllysine-residue (2-methylpropanoyl)transferase] S-(3-methylbutanoyl)dihydrolipoyllysine + CO2
show the reaction diagram
-
-
-
-
r
acyl-E1b-thiamine diphosphate + lipoyl-[lipoic acid-bearing domain]
2-oxo-acyl-S-lipoyl-[lipoic acid-bearing domain] + E1b-thiamine diphosphate
show the reaction diagram
-
reductive acylation of lipoyl-LBD
-
-
?
alpha-keto-beta-methylvaleric acid + NADH
?
show the reaction diagram
-
-
-
-
?
alpha-ketoisocaproic acid + NADH
?
show the reaction diagram
-
-
-
-
?
alpha-ketoisovaleric acid + NADH
?
show the reaction diagram
-
-
-
-
?
E1b-thiamine diphosphate + 2-oxo-acid
E1b-thiamine diphosphate-acyl + CO2
show the reaction diagram
-
decarboxylation, His146beta' and His291alpha are involved
-
-
?
pyruvate + NAD+ + CoA
acetyl-CoA + 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
2-oxo-isocaproate + NAD+ + CoA
3-methylbutanoyl-CoA + CO2 + NADH + H+
show the reaction diagram
3-methyl-2-oxobutanoate + NAD+ + CoA
2-methylpropanoyl-CoA + CO2 + NADH + H+
show the reaction diagram
-
-
-
-
?
3-methyl-2-oxobutanoate + [dihydrolipoyllysine-residue (2-methylpropanoyl)transferase] lipoyllysine
[dihydrolipoyllysine-residue (2-methylpropanoyl)transferase] S-(2-methylpropanoyl)dihydrolipoyllysine + CO2
show the reaction diagram
-
-
-
-
?
3-methyl-2-oxobutanoate + [dihydrolipoyllysine-residue(2-methylpropanoyl)transferase]-lipoyllysine
[dihydrolipoyllysine-residue (2-methylpropanoyl)transferase] S-(2-methylpropanoyl)dihydrolipoyllysine + CO2
show the reaction diagram
alpha-keto-beta-methylvaleric acid + NADH
?
show the reaction diagram
-
-
-
-
?
alpha-ketoisocaproic acid + NADH
?
show the reaction diagram
-
-
-
-
?
alpha-ketoisovaleric acid + NADH
?
show the reaction diagram
-
-
-
-
?
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
thiamine diphosphate
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
-
required
phosphate
-
required
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-chloro-4-methylpentanoate
-
inhibits component E1, modifies a histidine side chain
2-chloroisocaproate
-
-
2-oxobutanoate
-
-
2-oxoglutarate
-
-
2-Oxopentanoate
-
-
3-methyl-2-oxobutanoate
-
substrate inhibition
3-methyl-2-oxopentanoate
-
-
3-methylbutanoyl-CoA
4-(2-Thienyl)-2-oxo-3-butenoate
-
-
4-(3-Thienyl)-2-oxo-3-butenoate
4-methyl-2-oxopentanoate
alpha-Ketoisocaproate
-
allosteric inhibition, also by other branched-chain keto acids, although they are less effective as compared to alpha-ketoisocaproate
cinnamylpyruvate
Clofibrate
-
-
D-3-methyl-2-oxopentanoate
-
substrate inhibition
Furfurylidenepyruvate
isobutanoyl-CoA
-
0.2 mM, 48% inhibition
isopentanoyl-CoA
L-3-Methyl-2-oxopentanoate
-
substrate inhibition
microRNA 29a
-
total BCKD activity decreases over a 6-day period following a single transfection
-
microRNA 29b
-
total BCKD activity decreases over a 6-day period following a single transfection, is targeted to the mRNA for the dihydrolipoamide branched chain acyltransferase component of BCKD and prevents translation when bound
-
Skeletal muscle factor
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activity from liver, kidney and brains
-
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Activator protein
-
a protein in rat liver and kidney mitochondria that reactivates phosphorylated branched-chain complex without dephosphorylation
-
alpha-Chloroisocaproate
alpha-Ketoisocaproate
alpha-ketoisovalerate
-
activates through release of branched-chain alpha-keto acid dehydrogenase kinase from the BCKD complex
bezafibrate
-
i.e. 2-[4-[2-(4-chlorobenzamido)ethyl]phenoxy]-2-methylpropanoic acid. In rats treated with 5, 10, 20 mg/kg bezafibrate mean enzyme actual activity is 1.9, 2.9, and 4.3fold higher than in the control group, respectively. Enzymatic total activity increases by 1.3 and 1.6fold in rats treated with 10 and 20 mg/kg bezafibrate
Lipoamide
Skeletal muscle factor
-
stimulates enzyme from liver
-
thiamine
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thiamine increases the specific activity of the human liver enzyme complex
thiamine diphosphate
tumor necrosis factor-alpha
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0133
2-oxo-4-methylthiobutanoate
-
-
0.028 - 4.38
2-oxo-isovalerate
0.0075 - 0.056
2-oxobutanoate
0.29 - 2.5
2-oxoglutarate
0.00067 - 0.568
3-methyl-2-oxobutanoate
0.01 - 0.25
3-methyl-2-oxopentanoate
0.0062 - 0.2
4-methyl-2-oxopentanoate
0.033 - 0.06
coenzyme A
0.0105 - 0.0172
DL-3-methyl-2-oxopentanoate
0.03 - 0.12
NAD+
0.173 - 2.2
pyruvate
0.00055 - 0.04
thiamine diphosphate
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.195 - 9.4
2-oxo-isovalerate
0.1983 - 2.85
alpha-ketoisovaleric acid
0.197 - 6.38
thiamine diphosphate
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.15
4-(3-Thienyl)-2-oxo-3-butenoate
-
-
0.5
cinnamylpyruvate
-
-
0.0005
Furfurylidenepyruvate
-
-
0.0082 - 0.0136
isovaleryl-CoA
0.0177 - 0.023
NADH
additional information
additional information
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0582
-
-
3.13
purified branched-chain alpha-keto acid dehydrogenase multienzyme complex, substrate 2-oxo-isovalerate
5.5
-
multienzyme complex
8.8
-
multienzyme complex
38
-
E1 component expressed in Escherichia coli
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.8 - 7.4
-
-
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
22
-
assay at
30
-
assay at
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
fetal and maternal
Manually annotated by BRENDA team
-
activity increases when plants are placed in the dark, expression is strongly associated with the progression of leaf senescence
Manually annotated by BRENDA team
-
fetal and maternal
Manually annotated by BRENDA team
additional information
-
not detected in astrocytes and Bergmann glia cells
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
Thermoplasma acidophilum (strain ATCC 25905 / DSM 1728 / JCM 9062 / NBRC 15155 / AMRC-C165)
Thermoplasma acidophilum (strain ATCC 25905 / DSM 1728 / JCM 9062 / NBRC 15155 / AMRC-C165)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
46270
-
unlipoylated enzyme, mass spectrometric analysis
46460
-
lipoylated enzyme, mass spectrometric analysis
79400
-
gel filtration
154000
-
E1, heart
160000
-
wild-type enzyme and mutant enzymes S293E, S293A, S303A and S293E/S303E, gel filtration
167000
-
E1 component, equilibrium sedimentation
172000
-
equilibrium sedimentation
190000
additional information
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
24mer
-
E2 component
heterotetramer
-
x-ray crystallography
homotetramer
-
4 * 19850, gel filtration; 4 * 20000, SDS-PAGE
tetramer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phosphoprotein
-
branched-chain alpha-ketoacid dehydrogenase multienzyme complex BCKDC which is regulated by phosphorylation of component E1b, residue Ser292 of the alpha-domain, the phosphorylated recombinant E1b component shows highly reduced activity compared to the nonphosphorylated one
additional information
-
limited proteolytic analysis of E1b component, overview
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
20-25 mg/ml wild-type or recombinant His-tagged E1b component in 50 mM HEPES, pH 7.5, 0.25 M KCl, 0.5 mM PMSF, 1 mM benzamidine, 20 mM DTT, 5% v/v glycerol, vapour diffusion method, 20C, mixing with equal volume of well solution containing 1.4-1.6 M ammonium sulfate, 0.1 M sodium citrate, pH 5.8, 20 mM 2-mercaptoethanol, 4 mM MgCl2 or MnCl2, 4 mM thiamine diphosphate, maximal size after 10 day after microseeding, cryoprotection by well solution with 20% v/v glycerol, X-ray diffraction structure determination and analysis at 1.85-2.25 A resolution
-
crystals of E1b grown in the absence and presence of substrates at 22C via the vapor-diffusion method, key tyrosine residue in the E1b active site, functions as a conformational switch to reduce the reactivity of the thiamin diphosphate cofactor, the tyrosine switch further remodels an E1b loop region to promote E1b binding to E2b
-
purified wild-type and C-terminally His-tagged recombinant E1b component and mutants, 22C, hanging drop vapour diffusion method, Mg2+ or Mn2+, X-ray diffraction structure determination and analysis at 1.8 A resolution
-
crystallization of 4 forms of complex component E1: 1. E1 apoenzyme, 2. E1 holoenzyme, 3. E1 holoenzyme in complex with substrate analogue 4-methylpentanoate, 4. E1 holoenzyme in complex with substrate 4-methyl-2-oxopentanoate, hanging drop vapour diffusion method, 18C, 0.002 ml 10 mg/ml purified recombinant protein in 20 mM Tris-HCl, pH 8.0, 150 mM NaCl, 1 mM DTT, with equal volume of 0.002 ml of reservoir solution containing 0.7 M lithium sulfate, 60 mM sodium citrate, pH 5.6, against 0.4 ml reservoir solution, a few days, X-ray diffraction structure determination and analysis at 1.9-2.4 A resolution
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50
-
20 min, about 85% loss of activity without thiamin diphosphate. 20 min, about 65% loss of activity in presence of 0.2 mM thiamin diphosphate
additional information
-
thermal and cold denaturation kinetics of lipoic acid-bearing domain
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
maximum solubilization with minimum inactivation at 44 mM perchlorate carefully timed for 13 min
-
when saturated with thiamin diphosphate, the multienzyme complex is more stable to heat and chymotrypsin inactivation
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, after concentration and dialysis into 50 mM Tris-HCl buffer, 0.1 mM EDTA, 0.1 mM EGTA, 1 mM benzamidine, 1 mM PMSF, 1 mM DTT, pH 7.3, multienzyme complex is stable for at least 2 months
-
-70C, 10 mM Tris-HC1, pH 8.0, containing 0.01 mM EDTA, 0.2 M NaCI, 0.005% (v/v) 2-mercaptoethanol, and 50% (v/v) glycerol, 2 years, remains active
-
-70C, 50 mM potassium phosphate buffer, pH 7.4, 0.1 mM EDTA, 0.2 mM thiamine diphosphate, 0.2 mM phenylmethylsulfonyl fluoride, stable for at least 3 weeks
-
-70C, enzyme complex is stable for more than 8 months
-
-70C, multienzyme complex is stable for at least 2 years
-
0-5C, rapid loss of 20-30% of the activity, then stable for 3-4 months
-
0C, glycerol buffer, 20% loss of activity after 24 h
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
branched-chain alpha-keto acid dehydrogenase multienzyme complex BCKADH, consisting of 3 subunits/components E1-E3, from liver mitochondria, isolation of the complex components E1, E2, and E3
by Ni-NTA affinity chromatography
-
DEAE-Sepharose CL-6B column chromatography and heparin-Sepharose CL-6B column chromatography
-
E1 component, recombinant enzyme
-
enzyme complex components E1 and E2 from liver mitochondria
His Trap HP column chromatography and Strep-Tactin Superflow gravity column chromatography
-
His-Bind affinity chromatography and Q-Sepharose Hi-Trap column chromatography
-
hydrophobic interaction chromatography on phenyl-Sepharose; multienzyme complex
-
multienzyme complex
multienzyme complex; partial
-
partial
-
recombinant alpha- and beta-subunits of component E1 from Escherichia coli
-
recombinant C-terminally His-tagged lipoic acid-bearing domain LBD by nickel affinity chromatography
-
recombinant His-tagged lipoic acid-bearing domain of enzyme complex component E2 by nickel affinity chromatography and gel filtration to over 95% purity
-
recombinant lipoic acid-bearing domain of complex component E2 from Escherichia coli strain BL21(DE3) as His-tagged protein by nickel affinity chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
beta-subunit
DNA and amino acid sequence determination and analysis of E2
expressed in Escherichia coli BL-21 cells
-
expressed in Escherichia coli BL21(DE3) cells and BL21(DE3)pLysS cells
-
expressed in Escherichia coli strain DH5alpha
-
expression in Escherichia coli
-
expression in Pseudomonas putida and Escherichia coli; region containing the genes for the entire enzyme complex
-
expression of alpha- and beta-subunits of component E1 in Escherichia coli
-
expression of C-terminally His-tagged lipoic acid-bearing domain LBD, amino acid residues 1-84 of E2b
-
expression of C-terminally His-tagged lipoic acid-bearing domain of enzyme complex component E2 with an extra methionine at the N-terminus and a glutamic acid and leucine at the C-terminus in Escherichia coli strain BL21(DE3)
-
expression of E1 component in Escherichia coli as a homotetramer, in Escherichia coli only one translational start site for E1beta is recognized and only the 39000 Da product is produced, in addition the N-terminal amino acid of the 39000 Da E1beta subunit is Met instead of Leu
-
expression of N-and C-terminally His6-tagged component E1b and mutants, and of C-terminally His-tagged lipoic acid-bearing domain, amino acid residues 1-84 of component E2b, in vitro lipoylation of the latter
-
expression of the lipoic acid-bearing domain of complex component E2 in Escherichia coli strain BL21(DE3) as His-tagged protein
-
overexpression of bacterial branched-chain alpha-keto acid dehydrogenase multienzyme complex BCKDC, encoded by 4 genes, in Arabidopsis thaliana targeted to chloroplasts, about 7fold increased enzyme activity in the transgenic plants
-
region containing the genes for the entire enzyme complex
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
donor hepatocytes expressing wild type BCKDH engrafted into intermediate maple syrup urine disease recipient liver, increase liver BCKDH activity. Hepatocyte transplantation increases liver BCKDH activity 14.36% of control activity
-
in skeletal muscle, subunits E1alpha protein is significantly reduced in OLETF rats
-
mRNA expression is not upregulated by bezafibrate treatment
-
total BCKDC activity and protein abundance of E1alpha, E1beta, and E2 subunits are markedly lower in Otsuka Long-Evans Tokushima Fatty (OLETF) than in Long-Evans Tokushima Otsuka (LETO) rats at 8 and 25 weeks of age. In addition, hepatic BCKDC activity and protein amounts are significantly decreased in LETO rats aged 25 weeks than in LETO rats aged 8 weeks. In skeletal muscle, subunits E1beta and E2 proteins are significantly reduced in OLETF rats
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D108N
-
less thermostable than wild type enzyme
D200A
-
less thermostable than wild type enzyme
D200A/D108N
-
the mutant is defective in folding and assembly and as no detectable overall activity
D200N
-
less thermostable than wild type enzyme
D295A
-
site-directed mutagenesis of an alpha-subunit residue of component E1b, increased activity compared to the wild-type
E198Q/D200N
-
the mutant is defective in folding and assembly and as no detectable overall activity
F364C
-
E1alpha-missense mutation in type IA maple syrup urine disease that causes the occurence of exclusively alpha,beta dimers
H146A
-
site-directed mutagenesis, mutation of a beta'-subunit residue, no reductive acylation activity, low decarboxylation activity
H147N
-
site-directed mutagenesis, mutation of a beta'-subunit residue, no reductive acylation activity, low decarboxylation activity
H291A
-
site-directed mutagenesis, mutation of a alpha-subunit residue, highly reduced activity compared to the wild-type E1b component
H291N
-
site-directed mutagenesis, mutation of a alpha-subunit residue, highly reduced activity compared to the wild-type E1b component
H291Q
-
site-directed mutagenesis, mutation of a alpha-subunit residue, highly reduced activity compared to the wild-type E1b component
R287A
-
site-directed mutagenesis of an alpha-subunit residue of component E1b, highly increased Km and reduced activity compared to the wild-type
R301A
-
site-directed mutagenesis of an alpha-subunit residue of component E1b, increased Km and reduced activity compared to the wild-type
S292D
-
site-directed mutagenesis, mutation of a alpha-subunit residue of complex component E1b, reduced activity compared to the wild-type E1b component
S292E
-
site-directed mutagenesis, mutation of a alpha-subunit residue of complex component E1b, highly reduced activity compared to the wild-type E1b component
S292N
-
site-directed mutagenesis, mutation of a alpha-subunit residue of complex component E1b, reduced activity compared to the wild-type E1b component
S302A
-
site-directed mutagenesis, mutation of a alpha-subunit residue of complex component E1b, increased activity compared to the wild-type E1b component
T265R
-
missense mutation in type IA maple syrup urine disease that causes the occurence of both alpha2beta2 tetramers and lower molecular weight species
Y300A
-
site-directed mutagenesis of an alpha-subunit residue of component E1b, increased Km and reduced activity compared to the wild-type
Y300F
-
site-directed mutagenesis of an alpha-subunit residue of component E1b, increased Km and reduced activity compared to the wild-type
Y368C
-
missense mutation in type IA maple syrup urine disease that causes the occurence of both alpha2beta2 tetramers and lower molecular weight species
Y393N
-
E1alpha-missense mutation in type IA maple syrup urine disease that causes the occurence of exclusively alpha,beta dimers
S313A
-
twofold increase in Km-value but no change in turnover-number
S315A
-
mutation has no effect on Km-value or turnover-number
S319A
-
mutation has no effect on Km-value or turnover-number
D296A
-
inactive enzyme, no ability of component E1 apoenzyme to reconstitute with thiamine diphosphate
H292A
-
inactive enzyme, no binding of thiamine diphosphate
R288A
-
inactive enzyme, no detectable phosphorylation
S293E/S303E
-
mutation in the alpha-subunit, no activity
S303A
-
mutation in the alpha-subunit, no effect upon enzyme activity
S303E
-
mutation in the alpha-subunit, no effect upon enzyme activity
additional information
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
denaturating of the lipoic acid-bearing domain of enzyme complex component E2 by 8 M urea, refolding by 10fold dilution, folding kinetics of the reversible 2-step mechanism
-
isothermal urea denaturation and refolding kinetics of lipoic acid-bearing domain, 2-step mechanism, overview
-
reconstitution of the active multienzyme complex with resolved components
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
additional information
Show AA Sequence (1649 entries)
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