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show all sequences of 1.2.7.11

Crystal structures of archaeal 2-oxoacid:ferredoxin oxidoreductases from Sulfolobus tokodaii

Yan, Z.; Maruyama, A.; Arakawa, T.; Fushinobu, S.; Wakagi, T.; Sci. Rep. 6, 33061 (2016)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
expressed in Escherichia coli C43 (DE3) cells; expressed in Escherichia coli C43 (DE3) cells; expression in Escherichia coli (DE3); expression in Escherichia coli (DE3)
Sulfurisphaera tokodaii
Crystallization (Commentary)
Crystallization
Organism
crystals are grown at 20°C using sitting drop vapor diffusion. The structure of the recombinant enzyme StOFOR2 by the single-wavelength anomalous dispersion method is solved using a selenomethionine(SeMet)-labeled protein crystal, and the structures of the ligand-free (2.1 Å resolution) and pyruvate-complexed (2.2 Å) forms are determined. In the structure of the recombinant enzyme StOFOR2 in unreacted pyruvate complex form, the carboxylate group of pyruvate is recognized by Arg344 and Thr257 from the alpha-subunit. The binding pockets of the 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii surrounding the methyl or propyl group of the ligands are wider than that of 2-oxoacid oxidoreductase enzymes from Desulfovbrio africanus. A possible complex structural model is constructed by placing a Zn2+-containing dicluster ferredoxin of Sulfolobus tokodaii into the large pocket of the recombinant StOFOR2 enzyme, providing insight into the electron transfer between the two redox proteins; crystals of the StOFOR1 enzyme are prepared by co-crystallization with 50 mM 2-oxobutyrate and 1 mM CoA. Crystals are grown at 25°C using sitting drop vapor diffusion. In the structure of StOFOR1 co-crystallized with 2-oxobutyrate, electron density corresponding to a 1-hydroxypropyl group (post-decarboxylation state) is observed at the thiazole ring of thiamine diphosphate. The binding pockets of the 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii surrounding the methyl or propyl group of the ligands are wider than that of 2-oxoacid oxidoreductase enzymes from Desulfovbrio africanus; sitting drop vapor diffusion method, using 0.7 M ammonium tartrate dibasic and 0.1 M Tris-HCl (pH 8.5); sitting drop vapor diffusion method, using 15% (w/v) PEG3350, 0.1 M Tris-HCl (pH 8.5) and 0.1 M NaBr
Sulfurisphaera tokodaii
Engineering
Amino acid exchange
Commentary
Organism
D468A
inactive with 2-oxoglutarate and pyruvate as substrate; mutant enzyme StOFOR1 with mutation D468A in alpha-subunit. Vmax with pyruvate as substrate is 1.3% compared to wild-type enzyme. No activity is detected with 2-oxoglutarate
Sulfurisphaera tokodaii
K49I
inactive with 2-oxoglutarate as substrate; mutant enzyme StOFOR1 with mutation K49I in alpha-subunit. Vmax with pyruvate as substrate is 28% compared to wild-type enzyme, Km with pyruvate as substrate is 2.8fold higher as compared to wild-type enzyme. No activity is detected with 2-oxoglutarate
Sulfurisphaera tokodaii
S41A
mutant enzyme StOFOR1 with mutation S41A in alpha-subunit. Vmax with pyruvate as substrate is 29% compared to wild-type enzyme, Vmax with 2-oxoglutarate as substrate is 40% compared to wild-type enzyme, Km with pyruvate as substrate is 1.5fold higher as compared to wild-type enzyme, Km with 2-oxoglutarate as substrate is 1.5fold higher as compared to wild-type enzyme; the mutant shows reduced activity compared to the wild type enzyme
Sulfurisphaera tokodaii
T349L
mutant enzyme StOFOR1 with mutation T349L in alpha-subunit. Vmax with pyruvate as substrate is 43% compared to wild-type enzyme, Vmax with 2-oxoglutarate as substrate is 74% compared to wild-type enzyme, Km with pyruvate as substrate is 1.6fold higher as compared to wild-type enzyme, Km with 2-oxoglutarate as substrate is 1.1fold higher as compared to wild-type enzyme; the mutant shows reduced activity compared to the wild type enzyme
Sulfurisphaera tokodaii
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.32
-
pyruvate
pH 8.5, 80°C, cosubstrate: oxidized methyl viologen, enzyme StOFOR1; wild type enzyme, at pH 8.5 and 80°C
Sulfurisphaera tokodaii
0.49
-
pyruvate
mutant enzyme S41A, at pH 8.5 and 80°C; pH 8.5, 80°C, cosubstrate: oxidized methyl viologen, mutant enzyme StOFOR1 with mutation S41A in alpha-subunit
Sulfurisphaera tokodaii
0.51
-
pyruvate
mutant enzyme T349L, at pH 8.5 and 80°C; pH 8.5, 80°C, cosubstrate: oxidized methyl viologen, mutant enzyme StOFOR1 with mutation T349L in alpha-subunit
Sulfurisphaera tokodaii
0.91
-
pyruvate
mutant enzyme K49I, at pH 8.5 and 80°C; pH 8.5, 80°C, cosubstrate: oxidized methyl viologen, mutant enzyme StOFOR1 with mutation K49I in alpha-subunit
Sulfurisphaera tokodaii
1.6
-
pyruvate
pH 8.5, 80°C, cosubstrate: oxidized methyl viologen, enzyme StOFOR2; wild type enzyme, at pH 8.5 and 80°C
Sulfurisphaera tokodaii
2.1
-
2-oxoglutarate
pH 8.5, 80°C, cosubstrate: oxidized methyl viologen, enzyme StOFOR1; wild type enzyme, at pH 8.5 and 80°C
Sulfurisphaera tokodaii
2.3
-
2-oxoglutarate
mutant enzyme T349L, at pH 8.5 and 80°C; pH 8.5, 80°C, cosubstrate: oxidized methyl viologen, mutant enzyme StOFOR1 with mutation T349L in alpha-subunit
Sulfurisphaera tokodaii
3.2
-
2-oxoglutarate
mutant enzyme S41A, at pH 8.5 and 80°C; pH 8.5, 80°C, cosubstrate: oxidized methyl viologen, mutant enzyme StOFOR1 with mutation S41A in alpha-subunit
Sulfurisphaera tokodaii
15
-
2-oxoglutarate
pH 8.5, 80°C, cosubstrate: oxidized methyl viologen, enzyme StOFOR2; wild type enzyme, at pH 8.5 and 80°C
Sulfurisphaera tokodaii
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Mg2+
required; required
Sulfurisphaera tokodaii
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
34000
-
;
Sulfurisphaera tokodaii
70000
-
;
Sulfurisphaera tokodaii
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2-oxoglutarate + CoA + oxidized ferredoxin
Sulfurisphaera tokodaii
-
succinyl-CoA + CO2 + reduced ferredoxin + H+
-
-
?
2-oxoglutarate + CoA + oxidized ferredoxin
Sulfurisphaera tokodaii 7
-
succinyl-CoA + CO2 + reduced ferredoxin + H+
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Sulfurisphaera tokodaii
Q96XT2 and Q96XT4
Q96XT2: alpha-subunit (ST2435), Q96XT4: beta-subunit (ST2433); there is no evidence for the expression of the StOFOR2 (tk_24350/stk_24330 genes) in Sulfolobus tokodaii. Recombinant protein samples of STK_24350/STK_24330 are prepared and this enzyme is designated as StOFOR2
-
Sulfurisphaera tokodaii
Q96XT4
beta-subunit
-
Sulfurisphaera tokodaii
Q96Y66 and Q96Y68
Q96Y66: alpha-subunit (ST2300), Q96Y68: beta-subunit (ST2298)
-
Sulfurisphaera tokodaii
Q96Y68
beta-subunit
-
Sulfurisphaera tokodaii 7
Q96XT4
beta-subunit
-
Sulfurisphaera tokodaii 7
Q96Y68
beta-subunit
-
Sulfurisphaera tokodaii DSM 16993
Q96XT2 and Q96XT4
Q96XT2: alpha-subunit (ST2435), Q96XT4: beta-subunit (ST2433); there is no evidence for the expression of the StOFOR2 (tk_24350/stk_24330 genes) in Sulfolobus tokodaii. Recombinant protein samples of STK_24350/STK_24330 are prepared and this enzyme is designated as StOFOR2
-
Sulfurisphaera tokodaii DSM 16993
Q96Y66 and Q96Y68
Q96Y66: alpha-subunit (ST2300), Q96Y68: beta-subunit (ST2298)
-
Purification (Commentary)
Commentary
Organism
DEAE Sepharose column chromatography and Superdex 200 gel filtration; DEAE Sepharose column chromatography and Superdex 200 gel filtration
Sulfurisphaera tokodaii
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2-oxobutyrate + CoA + 2 oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii
propanoyl-CoA + CO2 + 2 reduced methyl viologen
-
-
-
?
2-oxobutyrate + CoA + 2 oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii 7
propanoyl-CoA + CO2 + 2 reduced methyl viologen
-
-
-
?
2-oxobutyrate + CoA + 2 oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii DSM 16993
propanoyl-CoA + CO2 + 2 reduced methyl viologen
-
-
-
?
2-oxoglutarate + CoA + 2 oxidized ferredoxin
because 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii lack the intramolecular Fd-like domain V, there is a large pocket surrounded by domains III and VI in each protomer, which appears to be able to bind an external ferredoxin molecule
739701
Sulfurisphaera tokodaii
succinyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
-
-
?
2-oxoglutarate + CoA + 2 oxidized ferredoxin
the recombinant enzyme StOFOR2 exhibits a preference for pyruvate over 2-oxoglutarate. Because 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii lack the intramolecular Fd-like domain V, there is a large pocket surrounded by domains III and VI in each protomer, which appears to be able to bind an external ferredoxin molecule
739701
Sulfurisphaera tokodaii
succinyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
-
-
?
2-oxoglutarate + CoA + 2 oxidized ferredoxin
the recombinant enzyme StOFOR2 exhibits a preference for pyruvate over 2-oxoglutarate. Because 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii lack the intramolecular Fd-like domain V, there is a large pocket surrounded by domains III and VI in each protomer, which appears to be able to bind an external ferredoxin molecule
739701
Sulfurisphaera tokodaii DSM 16993
succinyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
-
-
?
2-oxoglutarate + CoA + 2 oxidized ferredoxin
because 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii lack the intramolecular Fd-like domain V, there is a large pocket surrounded by domains III and VI in each protomer, which appears to be able to bind an external ferredoxin molecule
739701
Sulfurisphaera tokodaii DSM 16993
succinyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
-
-
?
2-oxoglutarate + CoA + oxidized ferredoxin
-
739701
Sulfurisphaera tokodaii
succinyl-CoA + CO2 + reduced ferredoxin + H+
-
-
-
?
2-oxoglutarate + CoA + oxidized ferredoxin
-
739701
Sulfurisphaera tokodaii 7
succinyl-CoA + CO2 + reduced ferredoxin + H+
-
-
-
?
2-oxoglutarate + CoA + oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii
succinyl-CoA + CO2 + reduced methyl viologen + H+
-
-
-
?
2-oxoglutarate + CoA + oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii 7
succinyl-CoA + CO2 + reduced methyl viologen + H+
-
-
-
?
pyruvate + CoA + 2 oxidized ferredoxin
because 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii lack the intramolecular Fd-like domain V, there is a large pocket surrounded by domains III and VI in each protomer, which appears to be able to bind an external ferredoxin molecule
739701
Sulfurisphaera tokodaii
acetyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
-
-
?
pyruvate + CoA + 2 oxidized ferredoxin
the recombinant enzyme StOFOR2 exhibits a preference for pyruvate over 2-oxoglutarate. The carboxylate group of pyruvate is recognized by Arg344 and Thr257 from the alpha-subunit of
739701
Sulfurisphaera tokodaii
acetyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
-
-
?
pyruvate + CoA + 2 oxidized ferredoxin
the recombinant enzyme StOFOR2 exhibits a preference for pyruvate over 2-oxoglutarate. The carboxylate group of pyruvate is recognized by Arg344 and Thr257 from the alpha-subunit of
739701
Sulfurisphaera tokodaii DSM 16993
acetyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
-
-
?
pyruvate + CoA + 2 oxidized ferredoxin
because 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii lack the intramolecular Fd-like domain V, there is a large pocket surrounded by domains III and VI in each protomer, which appears to be able to bind an external ferredoxin molecule
739701
Sulfurisphaera tokodaii DSM 16993
acetyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
-
-
?
pyruvate + CoA + 2 oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii
acetyl-CoA + CO2 + 2 reduced methyl viologen + 2 H+
-
-
-
?
pyruvate + CoA + 2 oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii DSM 16993
acetyl-CoA + CO2 + 2 reduced methyl viologen + 2 H+
-
-
-
?
pyruvate + CoA + oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii
?
-
-
-
?
pyruvate + CoA + oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii 7
?
-
-
-
?
Subunits
Subunits
Commentary
Organism
heterodimer
1 * 70000 + 1 * 34000, SDS-PAGE; 1 * 70000 + 1 * 34000, SDS-PAGE; crystals consist of two protomers (1 and 2), each of which consists of one heterodimer of alpha- and beta-subunits; crystals consist of two protomers (1 and 2), each of which consists of one heterodimer of alpha- and beta-subunits
Sulfurisphaera tokodaii
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
80
-
assay at; assay at
Sulfurisphaera tokodaii
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8.5
-
assay at; assay at
Sulfurisphaera tokodaii
Cofactor
Cofactor
Commentary
Organism
Structure
Ferredoxin
alpha/beta-subunit heterodimers lack an intramolecular ferredoxin domain. Because 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii lack the intramolecular Fd-like domain V, there is a large pocket surrounded by domains III and VI in each protomer, which appears to be able to bind an external ferredoxin molecule; alpha/beta-subunit heterodimers lack an intramolecular ferredoxin domain. Because 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii lack the intramolecular Fd-like domain V, there is a large pocket surrounded by domains III and VI in each protomer, which appears to be able to bind an external ferredoxin molecule
Sulfurisphaera tokodaii
thiamine diphosphate
alpha/beta-subunit heterodimers contain thiamin diphosphate; alpha/beta-subunit heterodimers contain thiamin diphosphate
Sulfurisphaera tokodaii
[4Fe-4S]-center
alpha/beta-subunit heterodimers contain 4Fe-4S cluster; alpha/beta-subunit heterodimers contain 4Fe-4S cluster
Sulfurisphaera tokodaii
Cloned(Commentary) (protein specific)
Commentary
Organism
expressed in Escherichia coli C43 (DE3) cells
Sulfurisphaera tokodaii
expression in Escherichia coli (DE3)
Sulfurisphaera tokodaii
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
Ferredoxin
alpha/beta-subunit heterodimers lack an intramolecular ferredoxin domain. Because 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii lack the intramolecular Fd-like domain V, there is a large pocket surrounded by domains III and VI in each protomer, which appears to be able to bind an external ferredoxin molecule
Sulfurisphaera tokodaii
thiamine diphosphate
alpha/beta-subunit heterodimers contain thiamin diphosphate
Sulfurisphaera tokodaii
thiamine diphosphate
-
Sulfurisphaera tokodaii
[4Fe-4S]-center
alpha/beta-subunit heterodimers contain 4Fe-4S cluster
Sulfurisphaera tokodaii
[4Fe-4S]-center
-
Sulfurisphaera tokodaii
Crystallization (Commentary) (protein specific)
Crystallization
Organism
crystals are grown at 20°C using sitting drop vapor diffusion. The structure of the recombinant enzyme StOFOR2 by the single-wavelength anomalous dispersion method is solved using a selenomethionine(SeMet)-labeled protein crystal, and the structures of the ligand-free (2.1 Å resolution) and pyruvate-complexed (2.2 Å) forms are determined. In the structure of the recombinant enzyme StOFOR2 in unreacted pyruvate complex form, the carboxylate group of pyruvate is recognized by Arg344 and Thr257 from the alpha-subunit. The binding pockets of the 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii surrounding the methyl or propyl group of the ligands are wider than that of 2-oxoacid oxidoreductase enzymes from Desulfovbrio africanus. A possible complex structural model is constructed by placing a Zn2+-containing dicluster ferredoxin of Sulfolobus tokodaii into the large pocket of the recombinant StOFOR2 enzyme, providing insight into the electron transfer between the two redox proteins
Sulfurisphaera tokodaii
crystals of the StOFOR1 enzyme are prepared by co-crystallization with 50 mM 2-oxobutyrate and 1 mM CoA. Crystals are grown at 25°C using sitting drop vapor diffusion. In the structure of StOFOR1 co-crystallized with 2-oxobutyrate, electron density corresponding to a 1-hydroxypropyl group (post-decarboxylation state) is observed at the thiazole ring of thiamine diphosphate. The binding pockets of the 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii surrounding the methyl or propyl group of the ligands are wider than that of 2-oxoacid oxidoreductase enzymes from Desulfovbrio africanus
Sulfurisphaera tokodaii
sitting drop vapor diffusion method, using 0.7 M ammonium tartrate dibasic and 0.1 M Tris-HCl (pH 8.5)
Sulfurisphaera tokodaii
sitting drop vapor diffusion method, using 15% (w/v) PEG3350, 0.1 M Tris-HCl (pH 8.5) and 0.1 M NaBr
Sulfurisphaera tokodaii
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
D468A
mutant enzyme StOFOR1 with mutation D468A in alpha-subunit. Vmax with pyruvate as substrate is 1.3% compared to wild-type enzyme. No activity is detected with 2-oxoglutarate
Sulfurisphaera tokodaii
D468A
inactive with 2-oxoglutarate and pyruvate as substrate
Sulfurisphaera tokodaii
K49I
mutant enzyme StOFOR1 with mutation K49I in alpha-subunit. Vmax with pyruvate as substrate is 28% compared to wild-type enzyme, Km with pyruvate as substrate is 2.8fold higher as compared to wild-type enzyme. No activity is detected with 2-oxoglutarate
Sulfurisphaera tokodaii
K49I
inactive with 2-oxoglutarate as substrate
Sulfurisphaera tokodaii
S41A
mutant enzyme StOFOR1 with mutation S41A in alpha-subunit. Vmax with pyruvate as substrate is 29% compared to wild-type enzyme, Vmax with 2-oxoglutarate as substrate is 40% compared to wild-type enzyme, Km with pyruvate as substrate is 1.5fold higher as compared to wild-type enzyme, Km with 2-oxoglutarate as substrate is 1.5fold higher as compared to wild-type enzyme
Sulfurisphaera tokodaii
S41A
the mutant shows reduced activity compared to the wild type enzyme
Sulfurisphaera tokodaii
T349L
mutant enzyme StOFOR1 with mutation T349L in alpha-subunit. Vmax with pyruvate as substrate is 43% compared to wild-type enzyme, Vmax with 2-oxoglutarate as substrate is 74% compared to wild-type enzyme, Km with pyruvate as substrate is 1.6fold higher as compared to wild-type enzyme, Km with 2-oxoglutarate as substrate is 1.1fold higher as compared to wild-type enzyme
Sulfurisphaera tokodaii
T349L
the mutant shows reduced activity compared to the wild type enzyme
Sulfurisphaera tokodaii
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.32
-
pyruvate
pH 8.5, 80°C, cosubstrate: oxidized methyl viologen, enzyme StOFOR1
Sulfurisphaera tokodaii
0.32
-
pyruvate
wild type enzyme, at pH 8.5 and 80°C
Sulfurisphaera tokodaii
0.49
-
pyruvate
pH 8.5, 80°C, cosubstrate: oxidized methyl viologen, mutant enzyme StOFOR1 with mutation S41A in alpha-subunit
Sulfurisphaera tokodaii
0.49
-
pyruvate
mutant enzyme S41A, at pH 8.5 and 80°C
Sulfurisphaera tokodaii
0.51
-
pyruvate
pH 8.5, 80°C, cosubstrate: oxidized methyl viologen, mutant enzyme StOFOR1 with mutation T349L in alpha-subunit
Sulfurisphaera tokodaii
0.51
-
pyruvate
mutant enzyme T349L, at pH 8.5 and 80°C
Sulfurisphaera tokodaii
0.91
-
pyruvate
pH 8.5, 80°C, cosubstrate: oxidized methyl viologen, mutant enzyme StOFOR1 with mutation K49I in alpha-subunit
Sulfurisphaera tokodaii
0.91
-
pyruvate
mutant enzyme K49I, at pH 8.5 and 80°C
Sulfurisphaera tokodaii
1.6
-
pyruvate
pH 8.5, 80°C, cosubstrate: oxidized methyl viologen, enzyme StOFOR2
Sulfurisphaera tokodaii
1.6
-
pyruvate
wild type enzyme, at pH 8.5 and 80°C
Sulfurisphaera tokodaii
2.1
-
2-oxoglutarate
pH 8.5, 80°C, cosubstrate: oxidized methyl viologen, enzyme StOFOR1
Sulfurisphaera tokodaii
2.1
-
2-oxoglutarate
wild type enzyme, at pH 8.5 and 80°C
Sulfurisphaera tokodaii
2.3
-
2-oxoglutarate
pH 8.5, 80°C, cosubstrate: oxidized methyl viologen, mutant enzyme StOFOR1 with mutation T349L in alpha-subunit
Sulfurisphaera tokodaii
2.3
-
2-oxoglutarate
mutant enzyme T349L, at pH 8.5 and 80°C
Sulfurisphaera tokodaii
3.2
-
2-oxoglutarate
pH 8.5, 80°C, cosubstrate: oxidized methyl viologen, mutant enzyme StOFOR1 with mutation S41A in alpha-subunit
Sulfurisphaera tokodaii
3.2
-
2-oxoglutarate
mutant enzyme S41A, at pH 8.5 and 80°C
Sulfurisphaera tokodaii
15
-
2-oxoglutarate
pH 8.5, 80°C, cosubstrate: oxidized methyl viologen, enzyme StOFOR2
Sulfurisphaera tokodaii
15
-
2-oxoglutarate
wild type enzyme, at pH 8.5 and 80°C
Sulfurisphaera tokodaii
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Mg2+
required
Sulfurisphaera tokodaii
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
34000
-
-
Sulfurisphaera tokodaii
70000
-
-
Sulfurisphaera tokodaii
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2-oxoglutarate + CoA + oxidized ferredoxin
Sulfurisphaera tokodaii
-
succinyl-CoA + CO2 + reduced ferredoxin + H+
-
-
?
2-oxoglutarate + CoA + oxidized ferredoxin
Sulfurisphaera tokodaii 7
-
succinyl-CoA + CO2 + reduced ferredoxin + H+
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
-
Sulfurisphaera tokodaii
DEAE Sepharose column chromatography and Superdex 200 gel filtration
Sulfurisphaera tokodaii
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2-oxobutyrate + CoA + 2 oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii
propanoyl-CoA + CO2 + 2 reduced methyl viologen
-
-
-
?
2-oxobutyrate + CoA + 2 oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii 7
propanoyl-CoA + CO2 + 2 reduced methyl viologen
-
-
-
?
2-oxobutyrate + CoA + 2 oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii DSM 16993
propanoyl-CoA + CO2 + 2 reduced methyl viologen
-
-
-
?
2-oxoglutarate + CoA + 2 oxidized ferredoxin
because 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii lack the intramolecular Fd-like domain V, there is a large pocket surrounded by domains III and VI in each protomer, which appears to be able to bind an external ferredoxin molecule
739701
Sulfurisphaera tokodaii
succinyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
-
-
?
2-oxoglutarate + CoA + 2 oxidized ferredoxin
the recombinant enzyme StOFOR2 exhibits a preference for pyruvate over 2-oxoglutarate. Because 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii lack the intramolecular Fd-like domain V, there is a large pocket surrounded by domains III and VI in each protomer, which appears to be able to bind an external ferredoxin molecule
739701
Sulfurisphaera tokodaii
succinyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
-
-
?
2-oxoglutarate + CoA + 2 oxidized ferredoxin
the recombinant enzyme StOFOR2 exhibits a preference for pyruvate over 2-oxoglutarate. Because 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii lack the intramolecular Fd-like domain V, there is a large pocket surrounded by domains III and VI in each protomer, which appears to be able to bind an external ferredoxin molecule
739701
Sulfurisphaera tokodaii DSM 16993
succinyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
-
-
?
2-oxoglutarate + CoA + 2 oxidized ferredoxin
because 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii lack the intramolecular Fd-like domain V, there is a large pocket surrounded by domains III and VI in each protomer, which appears to be able to bind an external ferredoxin molecule
739701
Sulfurisphaera tokodaii DSM 16993
succinyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
-
-
?
2-oxoglutarate + CoA + oxidized ferredoxin
-
739701
Sulfurisphaera tokodaii
succinyl-CoA + CO2 + reduced ferredoxin + H+
-
-
-
?
2-oxoglutarate + CoA + oxidized ferredoxin
-
739701
Sulfurisphaera tokodaii 7
succinyl-CoA + CO2 + reduced ferredoxin + H+
-
-
-
?
2-oxoglutarate + CoA + oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii
succinyl-CoA + CO2 + reduced methyl viologen + H+
-
-
-
?
2-oxoglutarate + CoA + oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii 7
succinyl-CoA + CO2 + reduced methyl viologen + H+
-
-
-
?
pyruvate + CoA + 2 oxidized ferredoxin
because 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii lack the intramolecular Fd-like domain V, there is a large pocket surrounded by domains III and VI in each protomer, which appears to be able to bind an external ferredoxin molecule
739701
Sulfurisphaera tokodaii
acetyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
-
-
?
pyruvate + CoA + 2 oxidized ferredoxin
the recombinant enzyme StOFOR2 exhibits a preference for pyruvate over 2-oxoglutarate. The carboxylate group of pyruvate is recognized by Arg344 and Thr257 from the alpha-subunit of
739701
Sulfurisphaera tokodaii
acetyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
-
-
?
pyruvate + CoA + 2 oxidized ferredoxin
the recombinant enzyme StOFOR2 exhibits a preference for pyruvate over 2-oxoglutarate. The carboxylate group of pyruvate is recognized by Arg344 and Thr257 from the alpha-subunit of
739701
Sulfurisphaera tokodaii DSM 16993
acetyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
-
-
?
pyruvate + CoA + 2 oxidized ferredoxin
because 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii lack the intramolecular Fd-like domain V, there is a large pocket surrounded by domains III and VI in each protomer, which appears to be able to bind an external ferredoxin molecule
739701
Sulfurisphaera tokodaii DSM 16993
acetyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
-
-
?
pyruvate + CoA + 2 oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii
acetyl-CoA + CO2 + 2 reduced methyl viologen + 2 H+
-
-
-
?
pyruvate + CoA + 2 oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii DSM 16993
acetyl-CoA + CO2 + 2 reduced methyl viologen + 2 H+
-
-
-
?
pyruvate + CoA + oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii
?
-
-
-
?
pyruvate + CoA + oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii 7
?
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
heterodimer
crystals consist of two protomers (1 and 2), each of which consists of one heterodimer of alpha- and beta-subunits
Sulfurisphaera tokodaii
heterodimer
1 * 70000 + 1 * 34000, SDS-PAGE
Sulfurisphaera tokodaii
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
80
-
assay at
Sulfurisphaera tokodaii
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8.5
-
assay at
Sulfurisphaera tokodaii
General Information
General Information
Commentary
Organism
metabolism
can utilize both pyruvate and 2-oxoglutarate, playing a key role in the central metabolism; there is no evidence for the expression of the StOFOR2 (tk_24350/stk_24330 genes) in Sulfolobus tokodaii. It may be expressed in vivo under some culture conditions
Sulfurisphaera tokodaii
General Information (protein specific)
General Information
Commentary
Organism
metabolism
can utilize both pyruvate and 2-oxoglutarate, playing a key role in the central metabolism
Sulfurisphaera tokodaii
metabolism
there is no evidence for the expression of the StOFOR2 (tk_24350/stk_24330 genes) in Sulfolobus tokodaii. It may be expressed in vivo under some culture conditions
Sulfurisphaera tokodaii
Other publictions for EC 1.2.7.11
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
739701
Yan
Crystal structures of archaeal ...
Sulfurisphaera tokodaii, Sulfurisphaera tokodaii 7, Sulfurisphaera tokodaii DSM 16993
Sci. Rep.
6
33061
2016
-
-
1
1
4
-
-
9
-
1
2
4
-
11
-
-
1
-
-
-
-
-
32
1
1
-
-
-
1
-
-
3
-
-
-
-
-
4
10
4
8
-
-
-
-
18
-
2
4
4
-
-
-
4
-
-
-
-
32
4
2
-
-
-
2
-
-
-
-
1
2
-
-
-
740343
Gibson
A structural phylogeny for und ...
Desulfovibrio africanus, Saccharolobus solfataricus
Curr. Opin. Struct. Biol.
41
54-61
2016
-
-
-
-
-
-
-
-
-
-
-
2
-
2
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
740367
Li
-
The catalytic bias of 2-oxoaci ...
Desulfovibrio africanus, Hydrogenobacter thermophilus
Electrochim. Acta
199
349-356
2016
-
-
-
-
-
-
-
-
-
-
-
2
-
2
-
-
-
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-
-
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2
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2
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-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
724470
Yan
Four Cys residues in heterodim ...
Sulfurisphaera tokodaii
Biochim. Biophys. Acta
1844
736-743
2014
-
-
-
-
5
-
-
-
-
1
-
1
-
3
-
-
1
-
-
-
-
-
1
1
1
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
5
-
-
-
-
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-
1
-
1
-
-
-
1
-
-
-
-
1
1
1
-
-
-
1
-
-
-
-
-
-
-
-
-
740620
Yokooji
Genetic examination of initial ...
Thermococcus kodakarensis
J. Bacteriol.
195
1940-1948
2013
-
-
-
-
-
-
-
-
-
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-
1
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4
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1
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1
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1
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-
-
-
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-
-
-
-
718962
Luo
Identification of the lysine r ...
Sulfurisphaera tokodaii
Biochim. Biophys. Acta
1794
335-340
2009
-
-
1
-
2
-
1
3
-
-
-
-
-
3
-
-
1
-
-
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1
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1
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1
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1
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2
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1
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3
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-
1
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-
-
-
1
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
674385
Park
Purifications and characteriza ...
Saccharolobus solfataricus, Saccharolobus solfataricus DSM 1616, Saccharolobus solfataricus P1
J. Biochem. Mol. Biol.
39
46-54
2006
-
-
-
-
-
-
-
4
-
-
3
-
-
16
-
-
1
-
-
-
1
-
18
2
2
-
-
5
2
1
-
1
-
-
-
-
-
-
1
-
-
-
-
-
-
4
-
-
3
-
-
-
-
1
-
-
1
-
18
2
2
-
-
5
2
1
-
-
-
-
-
-
6
6
673610
Nishizawa
Gene expression and characteri ...
Aeropyrum pernix
FEBS Lett.
579
2319-2322
2005
-
-
1
-
-
-
1
2
-
-
5
1
-
2
-
-
1
-
-
-
-
-
12
1
2
1
1
-
1
1
-
-
-
-
-
-
-
1
-
-
-
-
-
1
-
2
-
-
5
1
-
-
-
1
-
-
-
-
12
1
2
1
1
-
1
1
-
-
-
-
-
-
-
-
654838
Fukuda
Substrate recognition by 2-oxo ...
Sulfolobus sp., Sulfolobus sp. 7
Biochim. Biophys. Acta
1597
74-80
2002
-
-
-
-
-
-
-
6
-
-
-
-
-
16
-
-
-
-
-
-
-
-
6
-
1
-
-
-
1
-
-
-
-
-
-
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-
-
-
-
-
-
-
-
-
6
-
-
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-
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-
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-
-
6
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
719399
Fukuda
Role of a highly conserved YPI ...
Sulfolobus sp., Sulfolobus sp. 7
Eur. J. Biochem.
268
5639-5646
2001
-
-
1
-
14
-
-
20
-
1
3
-
-
17
-
-
1
-
-
-
-
-
6
1
2
-
2
21
2
-
-
1
-
-
-
-
-
1
1
-
14
-
-
-
-
20
-
1
3
-
-
-
-
1
-
-
-
-
6
1
2
-
2
21
2
-
-
-
-
-
-
-
22
22
725876
Iwasaki
Ferredoxin and related enzymes ...
Sulfolobus sp., Sulfolobus sp. 7
Methods Enzymol.
334
3-22
2001
-
-
-
-
-
-
-
2
-
2
2
-
-
3
-
-
1
-
-
-
-
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6
1
1
-
-
-
1
-
-
2
-
-
-
-
-
-
2
-
-
-
-
-
-
2
-
2
2
-
-
-
-
1
-
-
-
-
6
1
1
-
-
-
1
-
-
-
-
-
-
-
-
-
288435
Zhang
2-Oxoacid:ferredoxin oxidoredu ...
Sulfolobus sp. 7, Sulfolobus sp.
J. Biochem.
120
587-599
1996
-
-
1
-
-
-
-
2
-
2
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9
1
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1
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-
-
2
6
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1
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-
1
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-
1
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1
1
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-
-
-
-
2
-
2
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1
-
1
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-
-
2
6
-
1
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-
1
-
-
-
-
-
-
-
-
-
725332
Iwasaki
Ferredoxin-dependent redox sys ...
Sulfolobus sp., Sulfolobus sp. 7
J. Biol. Chem.
270
17878-17883
1995
-
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16
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1
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1
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1
1
-
-
-
288417
Kerscher
Purification and properties of ...
Halobacterium salinarum
Eur. J. Biochem.
116
587-594
1981
-
-
-
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1
1
4
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1
3
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2
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1
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3
1
1
1
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1
1
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1
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1
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1
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1
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4
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1
3
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1
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3
1
1
1
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1
1
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-