BRENDA - Enzyme Database

Versatile architecture of a bacterial aconitase B and its catalytic performance in the sequential reaction coupled with isocitrate dehydrogenase

Tsuchiya, D.; Shimizu, N.; Tomita, M.; Biochim. Biophys. Acta 1784, 1847-1856 (2008)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
4.2.1.3
gene acnB, expression of wild-type enzyme and the fusion protein ICDH-AcnB in Escherichia coli strain BL21 (DE3)
Escherichia coli
Engineering
EC Number
Amino acid exchange
Commentary
Organism
4.2.1.3
additional information
construction of a fusion protein of aconitase B and isocitrate dehydrogenase, ICDH and AcnB, i.e. ICDH-AcnB, structure determination of ICDH-AcnB, overview
Escherichia coli
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
4.2.1.3
additional information
-
additional information
steady-state kinetics
Escherichia coli
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
4.2.1.3
Mg2+
-
Escherichia coli
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.1.1.42
46200
-
2 * 46200, estimated from amino acid sequence
Escherichia coli
1.1.1.42
85000
-
Guinier analysis
Escherichia coli
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
4.2.1.3
cis-aconitate + H2O
Escherichia coli
-
isocitrate
-
-
?
4.2.1.3
citrate
Escherichia coli
-
cis-aconitate + H2O
-
-
?
4.2.1.3
additional information
Escherichia coli
weak interactions, which affects structure and function of the proteins, of aconitase B and isocitrate dehydrogenase, overview. Two monomeric AcnB regions associate with the homodimeric ICDH region. The versatile architecture of AcnB may alter the metabolic process involving the Krebs cycle
?
-
-
-
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.1.1.42
Escherichia coli
P08200
-
-
4.2.1.3
Escherichia coli
P36683
-
-
Purification (Commentary)
EC Number
Commentary
Organism
1.1.1.42
His-tagged enzyme is purified by TALON resin column chromatography, ICDH fused maltose-binding protein is purified by amylose resin chromatography
Escherichia coli
4.2.1.3
recombinant enzymes from Escherichia coli strain BL21(DE3) by gel filtration
Escherichia coli
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.1.1.42
isocitrate + NADP+
-
696416
Escherichia coli
2-oxoglutarate + CO2 + NADPH
-
-
-
?
1.1.1.42
additional information
does not use citrate as substrate
696416
Escherichia coli
?
-
-
-
-
4.2.1.3
cis-aconitate + H2O
-
696416
Escherichia coli
isocitrate
-
-
-
?
4.2.1.3
citrate
-
696416
Escherichia coli
cis-aconitate + H2O
-
-
-
?
4.2.1.3
additional information
weak interactions, which affects structure and function of the proteins, of aconitase B and isocitrate dehydrogenase, overview. Two monomeric AcnB regions associate with the homodimeric ICDH region. The versatile architecture of AcnB may alter the metabolic process involving the Krebs cycle
696416
Escherichia coli
?
-
-
-
-
4.2.1.3
additional information
the active sites within ICDH-AcnB catalyze the three consecutive reactions, in which citrate is converted to 2-oxoglutarate, via cisaconitate and isocitrate
696416
Escherichia coli
?
-
-
-
-
Subunits
EC Number
Subunits
Commentary
Organism
1.1.1.42
homodimer
2 * 46200, estimated from amino acid sequence
Escherichia coli
4.2.1.3
homodimer
structural analysis by X-ray scattering and modelling of wild-type and mutant enzymes, overview
Escherichia coli
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
4.2.1.3
8
-
assay at
Escherichia coli
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.42
NADP+
-
Escherichia coli
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
4.2.1.3
gene acnB, expression of wild-type enzyme and the fusion protein ICDH-AcnB in Escherichia coli strain BL21 (DE3)
Escherichia coli
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.42
NADP+
-
Escherichia coli
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
4.2.1.3
additional information
construction of a fusion protein of aconitase B and isocitrate dehydrogenase, ICDH and AcnB, i.e. ICDH-AcnB, structure determination of ICDH-AcnB, overview
Escherichia coli
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
4.2.1.3
additional information
-
additional information
steady-state kinetics
Escherichia coli
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
4.2.1.3
Mg2+
-
Escherichia coli
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.1.1.42
46200
-
2 * 46200, estimated from amino acid sequence
Escherichia coli
1.1.1.42
85000
-
Guinier analysis
Escherichia coli
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
4.2.1.3
cis-aconitate + H2O
Escherichia coli
-
isocitrate
-
-
?
4.2.1.3
citrate
Escherichia coli
-
cis-aconitate + H2O
-
-
?
4.2.1.3
additional information
Escherichia coli
weak interactions, which affects structure and function of the proteins, of aconitase B and isocitrate dehydrogenase, overview. Two monomeric AcnB regions associate with the homodimeric ICDH region. The versatile architecture of AcnB may alter the metabolic process involving the Krebs cycle
?
-
-
-
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.42
His-tagged enzyme is purified by TALON resin column chromatography, ICDH fused maltose-binding protein is purified by amylose resin chromatography
Escherichia coli
4.2.1.3
recombinant enzymes from Escherichia coli strain BL21(DE3) by gel filtration
Escherichia coli
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.1.1.42
isocitrate + NADP+
-
696416
Escherichia coli
2-oxoglutarate + CO2 + NADPH
-
-
-
?
1.1.1.42
additional information
does not use citrate as substrate
696416
Escherichia coli
?
-
-
-
-
4.2.1.3
cis-aconitate + H2O
-
696416
Escherichia coli
isocitrate
-
-
-
?
4.2.1.3
citrate
-
696416
Escherichia coli
cis-aconitate + H2O
-
-
-
?
4.2.1.3
additional information
weak interactions, which affects structure and function of the proteins, of aconitase B and isocitrate dehydrogenase, overview. Two monomeric AcnB regions associate with the homodimeric ICDH region. The versatile architecture of AcnB may alter the metabolic process involving the Krebs cycle
696416
Escherichia coli
?
-
-
-
-
4.2.1.3
additional information
the active sites within ICDH-AcnB catalyze the three consecutive reactions, in which citrate is converted to 2-oxoglutarate, via cisaconitate and isocitrate
696416
Escherichia coli
?
-
-
-
-
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.1.1.42
homodimer
2 * 46200, estimated from amino acid sequence
Escherichia coli
4.2.1.3
homodimer
structural analysis by X-ray scattering and modelling of wild-type and mutant enzymes, overview
Escherichia coli
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
4.2.1.3
8
-
assay at
Escherichia coli