BRENDA - Enzyme Database

Fumarate reductase and succinate oxidase activity of Escherichia coli complex II homologs are perturbed differently by mutation of the flavin binding domain

Maklashina, E.; Iverson, T.M.; Sher, Y.; Kotlyar, V.; Andrell, J.; Mirza, O.; Hudson, J.M.; Armstrong, F.A.; Rothery, R.A.; Weiner, J.H.; Cecchini, G.; J. Biol. Chem. 281, 11357-11365 (2006)

Data extracted from this reference:

Crystallization (Commentary)
EC Number
Crystallization
Organism
1.3.5.4
hanging drop vapor diffusion method, x-ray structure of mutant E49Q
Escherichia coli
Engineering
EC Number
Amino acid exchange
Commentary
Organism
1.3.5.4
E49A
decrease in catalytic efficiency of both fumarate reduction and succinate oxidation
Escherichia coli
1.3.5.4
E49Q
decrease in catalytic efficiency of both fumarate reduction and succinate oxidation
Escherichia coli
1.3.5.4
additional information
investigation on the role of the amino acid side chain in enzymes with Glu/Gln/Ala substitutions at fumarate reductase FrdA Glu49 and succinate dehydrogenase SdhA, EC 1.5.3.1, Gln50. The mutant enzymes with Ala substitutions in either Frd or Sdh remain functionally similar to their wild type counterparts. There are, however, dramatic changes in the catalytic properties when Glu and Gln are exchanged for each other in Frd and Sdh. Both enzymes are more efficient succinate oxidases when Gln is in the target position and a better fumarate reductase when Glu is present. Structural and catalytic analyses of the FrdA E49Q and SdhA Q50E mutants suggest that coulombic effects and the electronic state of the FAD are critical in dictating the preferred directionality of the succinate/fumarate interconversions
Escherichia coli
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
1.3.5.4
malonate
-
Escherichia coli
1.3.5.4
oxaloacetate
-
Escherichia coli
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.3.5.4
0.02
-
fumarate
wild-type, pH 7.0, 30°C
Escherichia coli
1.3.5.4
0.03
-
fumarate
mutant E49A, pH 7.0, 30°C
Escherichia coli
1.3.5.4
0.11
-
succinate
mutant E49A, pH 7.0, 30°C
Escherichia coli
1.3.5.4
0.22
-
succinate
mutant E49A, pH 7.0, 30°C
Escherichia coli
1.3.5.4
0.55
-
succinate
wild-type, pH 7.0, 30°C
Escherichia coli
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.3.5.4
Escherichia coli
P00363
-
-
Purification (Commentary)
EC Number
Commentary
Organism
1.3.5.4
-
Escherichia coli
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.3.5.4
fumarate + menaquinol
-
674532
Escherichia coli
succinate + menaquinone
-
-
-
r
1.3.5.4
succinate + menaquinone
-
674532
Escherichia coli
fumarate + menaquinol
-
-
-
r
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.3.5.4
1
-
fumarate
mutant E49A, pH 7.0, 30°C
Escherichia coli
1.3.5.4
2.4
-
succinate
mutant E49A, pH 7.0, 30°C
Escherichia coli
1.3.5.4
4
-
succinate
mutant E49A, pH 7.0, 30°C
Escherichia coli
1.3.5.4
30
-
succinate
wild-type, pH 7.0, 30°C
Escherichia coli
1.3.5.4
32
-
fumarate
mutant E49A, pH 7.0, 30°C
Escherichia coli
1.3.5.4
250
-
fumarate
wild-type, pH 7.0, 30°C
Escherichia coli
Ki Value [mM]
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
1.3.5.4
0.00006
-
oxaloacetate
mutant E49A, pH 7.0, 30°C
Escherichia coli
1.3.5.4
0.0001
-
oxaloacetate
mutant E49A, pH 7.0, 30°C
Escherichia coli
1.3.5.4
0.0003
-
oxaloacetate
wild-type, pH 7.0, 30°C
Escherichia coli
1.3.5.4
0.005
-
malonate
mutant E49A, pH 7.0, 30°C
Escherichia coli
1.3.5.4
0.01
-
malonate
mutant E49A, pH 7.0, 30°C
Escherichia coli
1.3.5.4
0.025
-
malonate
wild-type, pH 7.0, 30°C
Escherichia coli
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
1.3.5.4
hanging drop vapor diffusion method, x-ray structure of mutant E49Q
Escherichia coli
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
1.3.5.4
E49A
decrease in catalytic efficiency of both fumarate reduction and succinate oxidation
Escherichia coli
1.3.5.4
E49Q
decrease in catalytic efficiency of both fumarate reduction and succinate oxidation
Escherichia coli
1.3.5.4
additional information
investigation on the role of the amino acid side chain in enzymes with Glu/Gln/Ala substitutions at fumarate reductase FrdA Glu49 and succinate dehydrogenase SdhA, EC 1.5.3.1, Gln50. The mutant enzymes with Ala substitutions in either Frd or Sdh remain functionally similar to their wild type counterparts. There are, however, dramatic changes in the catalytic properties when Glu and Gln are exchanged for each other in Frd and Sdh. Both enzymes are more efficient succinate oxidases when Gln is in the target position and a better fumarate reductase when Glu is present. Structural and catalytic analyses of the FrdA E49Q and SdhA Q50E mutants suggest that coulombic effects and the electronic state of the FAD are critical in dictating the preferred directionality of the succinate/fumarate interconversions
Escherichia coli
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
1.3.5.4
malonate
-
Escherichia coli
1.3.5.4
oxaloacetate
-
Escherichia coli
Ki Value [mM] (protein specific)
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
1.3.5.4
0.00006
-
oxaloacetate
mutant E49A, pH 7.0, 30°C
Escherichia coli
1.3.5.4
0.0001
-
oxaloacetate
mutant E49A, pH 7.0, 30°C
Escherichia coli
1.3.5.4
0.0003
-
oxaloacetate
wild-type, pH 7.0, 30°C
Escherichia coli
1.3.5.4
0.005
-
malonate
mutant E49A, pH 7.0, 30°C
Escherichia coli
1.3.5.4
0.01
-
malonate
mutant E49A, pH 7.0, 30°C
Escherichia coli
1.3.5.4
0.025
-
malonate
wild-type, pH 7.0, 30°C
Escherichia coli
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.3.5.4
0.02
-
fumarate
wild-type, pH 7.0, 30°C
Escherichia coli
1.3.5.4
0.03
-
fumarate
mutant E49A, pH 7.0, 30°C
Escherichia coli
1.3.5.4
0.11
-
succinate
mutant E49A, pH 7.0, 30°C
Escherichia coli
1.3.5.4
0.22
-
succinate
mutant E49A, pH 7.0, 30°C
Escherichia coli
1.3.5.4
0.55
-
succinate
wild-type, pH 7.0, 30°C
Escherichia coli
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.3.5.4
-
Escherichia coli
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.3.5.4
fumarate + menaquinol
-
674532
Escherichia coli
succinate + menaquinone
-
-
-
r
1.3.5.4
succinate + menaquinone
-
674532
Escherichia coli
fumarate + menaquinol
-
-
-
r
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.3.5.4
1
-
fumarate
mutant E49A, pH 7.0, 30°C
Escherichia coli
1.3.5.4
2.4
-
succinate
mutant E49A, pH 7.0, 30°C
Escherichia coli
1.3.5.4
4
-
succinate
mutant E49A, pH 7.0, 30°C
Escherichia coli
1.3.5.4
30
-
succinate
wild-type, pH 7.0, 30°C
Escherichia coli
1.3.5.4
32
-
fumarate
mutant E49A, pH 7.0, 30°C
Escherichia coli
1.3.5.4
250
-
fumarate
wild-type, pH 7.0, 30°C
Escherichia coli
KCat/KM [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.3.5.4
11
-
succinate
mutant E49A, pH 7.0, 30°C
Escherichia coli
1.3.5.4
36
-
succinate
mutant E49A, pH 7.0, 30°C
Escherichia coli
1.3.5.4
54
-
succinate
wild-type, pH 7.0, 30°C
Escherichia coli
1.3.5.4
97
-
fumarate
mutant E49A, pH 7.0, 30°C
Escherichia coli
1.3.5.4
232
-
fumarate
wild-type, pH 7.0, 30°C
Escherichia coli
KCat/KM [mM/s] (protein specific)
EC Number
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.3.5.4
11
-
succinate
mutant E49A, pH 7.0, 30°C
Escherichia coli
1.3.5.4
36
-
succinate
mutant E49A, pH 7.0, 30°C
Escherichia coli
1.3.5.4
54
-
succinate
wild-type, pH 7.0, 30°C
Escherichia coli
1.3.5.4
97
-
fumarate
mutant E49A, pH 7.0, 30°C
Escherichia coli
1.3.5.4
232
-
fumarate
wild-type, pH 7.0, 30°C
Escherichia coli