Information on EC 1.3.5.4 - fumarate reductase (quinol)

Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Mark a special word or phrase in this record:
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


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

EC NUMBER
COMMENTARY
1.3.5.4
-
RECOMMENDED NAME
GeneOntology No.
fumarate reductase (quinol)
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
succinate + a quinone = fumarate + a quinol
show the reaction diagram
-
-
-
-
succinate + a quinone = fumarate + a quinol
show the reaction diagram
reduction of menaquinone-7 is the rate-limiting step in succinate respiration
-
PATHWAY
KEGG Link
MetaCyc Link
Biosynthesis of secondary metabolites
-
incomplete reductive TCA cycle
-
Metabolic pathways
-
Microbial metabolism in diverse environments
-
mixed acid fermentation
-
NADH to fumarate electron transfer
-
pyruvate fermentation to propionate I
-
Pyruvate metabolism
-
reductive TCA cycle I
-
reductive TCA cycle II
-
respiration (anaerobic)
-
TCA cycle III (helicobacter)
-
TCA cycle VI (obligate autotrophs)
-
SYSTEMATIC NAME
IUBMB Comments
succinate:quinone oxidoreductase
The enzyme, which is found in anaerobic and facultative organisms such as bacteria, parasitic helminthes, and lower marine organisms, utilizes low potential quinols, such as menaquinol and rhodoquinol, to reduce fumarate as the final step of an anaerobic respiratory chain. The enzyme is known as complex II of the electron transfer chain, similarly to EC 1.3.5.1, succinate dehydrogenase (quinone), to which it is closely related.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
8-methylmenaquinol:fumarate reductase
-
-
FRD
-
-
-
-
menaquinol-fumarate oxidoreductase
-
-
-
-
menaquinol-fumarate oxidoreductase
P00363
cf. EC 1.3.5.1, succinate-ubiquinone oxidoreductase, structurally and functionally related membrane-bound enzyme complexes
menaquinol:fumarate oxidoreductase
-
-
methylmenaquinol:fumarate reductase
-
-
non-classical succinate:quinone reductase
-
-
QFR
P92507
-
quinol-fumarate reductase
P92507
-
quinol-fumarate reductase
-
-
quinol:fumarate reductase
-
-
succinate:menaquinone 7-reductase
-
-
succinate:menaquinone oxidoreductase
-
-
succinate:menaquinone reductase
-
-
succinate:MK reductase
-
-
succinate:quinone oxidoreductase
-
ambiguous
CAS REGISTRY NUMBER
COMMENTARY
9076-99-7
cf EC 1.3.1.6
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
physiological function
-
is involved in anaerobic respiration with fumarate as the terminal electron acceptor, and is part of an electron transport chain catalysing the oxidation of various donor substrates by fumarate
physiological function
-
the enzyme is involved in anaerobic respiration with fumarate as the terminal electron acceptor, and is part of an electron transport chain catalysing the oxidation of various donor substrates by fumarate
physiological function
-
the enzyme is involved in anaerobic metabolism
physiological function
P00363
the QFR complex provides electron transport during anaerobic cell growth conditions. The transcription of the frdABCD operon responds to environmental as well as internal cell signals to modulate gene expression. The transcription is coupled to that of the succinate-ubiquinone oxidase, EC 1.3.5.1, overview
physiological function
-
fumarate reduction by NADH is catalyzed by an electron transport chain consisting of NADH dehydrogenase NADH:menaquinone reductase, menaquinone, and succinate dehydrogenase operating in the reverse direction, i.e. menaquinol:fumarate reductase. In sdh or aro mutant strains, which lack succinate dehydrogenase or menaquinone, respectively, the activity of fumarate reduction by NADH is missing. The membrane fraction of a mutant lacking functional sdh genes catalyzes fumarate reduction by NADH or 2,3-dimethyl-1,4-naphthoquinol with less than 7% of the wild-type activities. In resting cells fumarate reduction requires glycerol or glucose as the electron donor, which presumably supply NADH for fumarate reduction
physiological function
-
enzyme belongs to a system of electron transport phosphorylation in which formate functions as the donor and fumarate as the terminal acceptor. Menaquinone is an obligatory redox mediator of formate-fumarate reductase electron transport phosphorylation system
physiological function
-
fumarate reductase, which is proficient in succinate oxidation, is able to functionally replace succinate-ubiquinone oxidoreductase in aerobic respiration when conditions are used to allow the expression of the frdABCD operon aerobically. Expression of plasmids which utilize the FRD promoter of the frdABCD operon fused to the sdhCDAB genes to drive expression shows that, under anaerobic growth conditions where fumarate is utilized as the terminal electron acceptor, succinate-ubiquinone oxidoreductase would function to support anaerobic growth of Escherichia coli
physiological function
-
the FrdD subunit has an essential role both in the interaction of the enzyme with reduced menaquinone and thus in anaerobic respiration with fumarate as electron acceptor, and in binding the enzyme to the membrane
physiological function
P92507
the enzyme is part of the complex II, which in the anaerobic respiratory chain of the parasitic nematode Ascaris suum, couples the reduction of fumarate to the oxidation of rhodoquinol. Critical role of the low redox potential of rhodoquinol in the fumarate reduction of Ascaris suum complex II
additional information
-
the E-pathway of transmembrane proton transfer is essential for catalysis by the diheme-containing quinol:fumarate reductase, molecular dynamics simulations, overview. The redox state of heme groups has a crucial effect on the connectivity patterns of mobile internal water molecules that can transiently support proton transfer from the bD-C-propionate to Glu-C180. The short H-bonding paths formed in the reduced states can lead to high proton conduction rates. The bD-C-propionate group is the branching point connecting proton transfer to the E-pathway from the quinol-oxidation site via interactions with the heme bD ligand His-C44, essential functional role of His-C44, hydrogen-bonded networks between the bD-C-propionate and Glu180, overview
additional information
P92507
enzyme structure-function relationship, overview
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2,3-dimethyl-1,4-naphthoquinol + fumarate
?
show the reaction diagram
-
-
-
-
?
fumarate + 2,3-dimethyl-1,4-naphthohydroquinone
succinate + 2,3-dimethyl-1,4-naphthoquinone
show the reaction diagram
-
-
-
?
fumarate + 2,3-dimethyl-1,4-naphthohydroquinone
succinate + 2,3-dimethyl-1,4-naphthoquinone
show the reaction diagram
-
-
-
?
fumarate + 2,3-dimethyl-1,4-naphthohydroquinone
succinate + 2,3-dimethyl-1,4-naphthoquinone
show the reaction diagram
-
reaction catalyzed by fumarate reductase complex, the site of the complex reacting with fumarate is situated on the 79000 Da subunit, and the site reacting with dimethylnaphthohydroquinone is cytochrome b
-
?, r
fumarate + 2,3-dimethyl-1,4-naphthohydroquinone
succinate + 2,3-dimethyl-1,4-naphthoquinone
show the reaction diagram
-
cytochrome b-dependent in both directions
-
r
fumarate + 2,3-dimethyl-1,4-naphthohydroquinone
succinate + 2,3-dimethyl-1,4-naphthoquinone
show the reaction diagram
-
mutation of His-82 to Arg in fumarate reductase subunit C prevents oxidation of 2,3-dimethyl-1,4-naphthohydroquinone
-
r
fumarate + 8-methylmenaquinol-6
succinate + 8-methylmenaquinone-6
show the reaction diagram
-
-
-
-
?
fumarate + a menaquinol
succinate + a menaquinone
show the reaction diagram
-
the enzyme is involved in anaerobic metabolism
-
-
?
fumarate + anthrahydroquinonesulfonate
succinate + anthraquinonesulfonate
show the reaction diagram
-
-
-
?
fumarate + anthrahydroquinonesulfonate
succinate + anthraquinonesulfonate
show the reaction diagram
-
reaction catalyzed in the presence or absence of cytochrome b
-
?
fumarate + menaquinol
succinate + ?
show the reaction diagram
-
-
-
?
fumarate + menaquinol
succinate + ?
show the reaction diagram
-
-
-
?
fumarate + menaquinol
succinate + menaquinone
show the reaction diagram
-
-
-
-
r
fumarate + menaquinol
succinate + menaquinone
show the reaction diagram
-
-
-
-
r
fumarate + menaquinol
succinate + menaquinone
show the reaction diagram
P00363
-
-
-
r
fumarate + menaquinol
succinate + menaquinone
show the reaction diagram
-
the enzyme catalyzes the terminal step of the phosphorylative electron transport
-
?
fumarate + menaquinol
succinate + menaquinone
show the reaction diagram
-
the enzyme catalyzes the terminal step of the phosphorylative electron transport
-
?
fumarate + menaquinol
succinate + menaquinone
show the reaction diagram
P00363
enzyme is expressed under anaerobic conditions, transcription is coupled to that of the succinate-ubiquinone oxidase, EC 1.3.5.1
-
-
r
succinate + a menaquinone
fumarate + a menaquinol
show the reaction diagram
-
-, the enzyme is involved in anaerobic respiration with fumarate as the terminal electron acceptor, and is part of an electron transport chain catalysing the oxidation of various donor substrates by fumarate
-
-
?
succinate + menaquinone
fumarate + menaquinol
show the reaction diagram
-
-
-
-
r
succinate + menaquinone
fumarate + menaquinol
show the reaction diagram
-
-
-
-
r
succinate + menaquinone
fumarate + menaquinol
show the reaction diagram
-
-
-
-
r
succinate + menaquinone
fumarate + menaquinol
show the reaction diagram
P00363
-
-
-
r
succinate + methylene blue
?
show the reaction diagram
-
-
-
-
?
succinate + phenazine methosulfate
fumarate + reduced phenazine methosulfate
show the reaction diagram
-
-
-
-
r
succinate + phenazine methosulfate + 2,6-dichlorophenolindophenol
fumarate + ?
show the reaction diagram
-
quinone reduction by Rhodothermus marinus succinate:menaquinone oxidoreductase is not stimulated by the membrane potential
-
-
?
succinate + rhodoquinone
fumarate + rhodoquinol
show the reaction diagram
P92507
-, rhodoquinone binding site, overview
-
-
?
fumarate + menaquinol-6
succinate + menaquinone-6
show the reaction diagram
-
-
-
-
r
additional information
?
-
-
the fumarate reductase complex has two different reactive sites, which are essential for its function in the phosphorylative electron transport of the bacterium
-
-
-
additional information
?
-
-
enzyme also accepts artificial electron acceptors, reaction of EC 1.3.99.1
-
-
-
additional information
?
-
-
enzyme also accepts reduced decylubiquinone, reaction of EC 1.3.5.1
-
-
-
additional information
?
-
-
enzyme operates with both natural quinones, ubiquinone and menaquinone, at a single quinone binding site. Residue Lys228 in subunit FrdB provides a strong hydrogen bond to menaquinone and is essential for reactions with both quinone types. There is similar hydrogen bonding of the C1 carbonyl of both MQ and UQ, whereas there is different hydrogen bonding for their C4 carbonyls
-
-
-
additional information
?
-
-
evidence for proton potential dependent catalysis of succinate oxidation by quinone as well as for proton potential generation upon catalysis of fumarate reduction by quinol
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
fumarate + 2,3-dimethyl-1,4-naphthohydroquinone
succinate + 2,3-dimethyl-1,4-naphthoquinone
show the reaction diagram
-
-
-
?
fumarate + a menaquinol
succinate + a menaquinone
show the reaction diagram
-
the enzyme is involved in anaerobic metabolism
-
-
?
fumarate + menaquinol
succinate + menaquinone
show the reaction diagram
-
the enzyme catalyzes the terminal step of the phosphorylative electron transport
-
?
succinate + a menaquinone
fumarate + a menaquinol
show the reaction diagram
-
the enzyme is involved in anaerobic respiration with fumarate as the terminal electron acceptor, and is part of an electron transport chain catalysing the oxidation of various donor substrates by fumarate
-
-
?
succinate + rhodoquinone
fumarate + rhodoquinol
show the reaction diagram
P92507
-
-
-
?
fumarate + menaquinol
succinate + menaquinone
show the reaction diagram
-
the enzyme catalyzes the terminal step of the phosphorylative electron transport
-
?
additional information
?
-
-
the fumarate reductase complex has two different reactive sites, which are essential for its function in the phosphorylative electron transport of the bacterium
-
-
-
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
cytochrome b
-
has a MW of 25000 Da, a midpoint potential of - 15 mV and is reducible by dimethylnaphthohydroquinone in the absence of the other subunits
-
cytochrome b
-
fumarate reductase contains a diheme cytochrome b
-
cytochrome b
-
2 mol per mol FAD; 2 mol per mol of FAD
-
cytochrome b
-
the enzyme contains a cytochrome b with a midpoint potential of -20 mV, referred to as the high-potential cytochrome b and a cytochrome b with a midpoint potential of -200 mV, referred to as the low-potential cytochrome b
-
cytochrome b
-
-
-
FAD
-
one protein-bound FAD linked to the 79000 Da peptide
FAD
-
covalently bound to flavoprotein subunit
FAD
-
subunit A comprises a large FAD-binding domain
FAD
-
non-covalenly bound. In the enzyme containing a mutant A86H flavoprotein subunit the FAD is covalently bound
FAD
P00363
covalently attached to the enzyme to enable succinate oxidation
FAD
-
stoichiometric ratio between covalently bound FAD and the iron-sulfur cluster is 1:1. Protoheme IX is present in about 2:1 stoichiometry to covalently bound FAD
FAD
-
prosthetic group of fumarate reductase is covalently bound FAD. The specific activity of fumarate reductase is increased to the same extent as the content of the covalently bound FAD when the membrane is fractionated with cholate and ammonium sulfate. The acid-extractable FAD is removed by this procedure
FAD
P92507
the FAD prosthetic group is held in the FAD binding domain by a covalent bond to His A79 and by hydrogen bonds with highly conserved residues, overview
heme
-
enzyme contains about 11 iron atoms per complex, which is expected if the enzyme contains one [2Fe-2S] cluster, one [3Fe-4S] cluster, one [4Fe-4S] cluster and two type b hemes. Protoheme IX is present in about 2:1 stoichiometry to covalently bound FAD
heme
-
a diheme-containing enzyme, each heterotrimer contains two heme b groups bound by the transmembrane subunit C, which are termed the proximal heme, bP, and the distal heme, bD, according to the relative proximity to the hydrophilic subunits A and B
heme
P92507
-
heme b
-
the enzyme contains one hydrophobic subunit (menaquinol-oxidising subunit C) with two haem b groups. The binding of the two heme molecules is described. The close proximity between the two hemes offers a straightforward possibility for transmembrane electron transfer
iron-sulfur centre
-
enzyme contains about 11 iron atoms per complex, which is expected if the enzyme contains one [2Fe-2S] cluster, one [3Fe-4S] cluster, one [4Fe-4S] cluster and two type b hemes. The purified mQFR complex has two iron-sulfur centers of the ferredoxin type that are paramagnetic in the reduced state, 2Fe-2S and 4Fe-4S, and one iron-sulfur center of the high potential type that is paramagnetic in the oxidized state, 3Fe-4S. Centers 2Fe-2S and 4Fe-4S exhibit a large difference in their redox midpoint potential, center 2Fe-2S is reducible with succinate, whereas the latter one can only be reduced by very low potential reductant such as dithionite
iron-sulfur centre
-
the iron-sulfur protein of the electron transport phosphorylation system is the donor for fumarate reductase
menaquinol-6
-
-
additional information
-
measurement of the redox potentials of the sulfur-centers
-
additional information
-
measurement of the redox potentials of the sulfur-centers; the enzyme contains iron-sulfur centers
-
additional information
-
the enzyme contains iron-sulfur centers
-
additional information
-
the enzyme contains iron-sulfur centers
-
additional information
-
two hydrophobic subunits (C and D) which bind either no haem b group
-
additional information
P92507
the enzyme structure comprises four subunits and five co-factors, cofactor structure comparisons, overview
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Fe
-
4Fe-4S, bound to Fp subunit or bridging between Ip and Fp subunits; non-heme iron, types of Fe-S clusters: 2Fe-2S, bound to Ip: iron-sulfur protein subunit, smaller subunit
Fe2+
P92507
of the three iron-sulfur centres bound to subunit Ip, [2Fe-2S] is coordinated by four cysteine residues,B89, B94, B97 and B109, and located in the N-terminal domain, whereas [4Fe-4S] and [3Fe-4S] that are coordinated by four, B182, B185, B188, and B249, and three, B192, B239 and B245, cysteine residues, respectively, are bound to the C-terminal domain. These iron-sulfur centres are also surrounded with highly conserved hydrophobic amino acid residues
Iron
-
attached to menaquinol-oxidising subunit C on the cytoplasmic side of the membrane is subunit B, containing the [3Fe-4S], [4Fe-4S], and [2Fe-2S] iron-sulphur centres (in the order of increasing distance from menaquinol-oxidising subunit C)
Iron
P00363
iron-sulfur subunit with 3 distinct [4Fe-4S], [3Fe-3S], and [2Fe-2S] clusters, i.e., organized in 2 domains, all participate in electron transfer, overview
Iron
-
enzyme contains about 11 iron atoms per complex, which is expected if the enzyme contains one [2Fe-2S] cluster, one [3Fe-4S] cluster, one [4Fe-4S] cluster and two type b hemes
Iron
-
the iron-sulfur protein of the electron transport phosphorylation system is the donor for fumarate reductase
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2-(n-heptyl)-4-hydroxy-quinoline N-oxide
-
-
2-heptyl-4-hydroxyquinoline N-oxide
-
inhibitor blocks the binding of menaquinol at the proximal quinone binding-site, crystallization studies
2-n-heptyl-4-hydroxyquinoline-N-oxide
-
site of inhibition is located at the oxidation side of cytochrome b
2-n-heptyl-4-hydroxyquinoline-N-oxide
-
-
2-[1-(p-chlorophenyl)ethyl] 4,6-dinitrophenol
-
inhibitor blocks the binding of menaquinol at the proximal quinone binding-site, crystallization studies
4-Chloromercuriphenyl sulfonate
-
inhibits the succinate oxidation by cell-derived particles
4-Chloromercuriphenylsulfonate
-
inhibits the oxidation of reduced menaquinone by fumarate. Fumarate reductase, measured with reduced benzylviologen as the donor, is not affected
antimycin A
-
site of inhibition is located at the oxidation side of cytochrome b
oxaloacetate
-
-
Pentachlorophenol
P00363
-
malonate
-
competitive
additional information
-
not inhibitory: heptyl 4-hydroxyquinoline N-oxide
-
additional information
-
not inhibitory: thenoyltrifluoroacetone, 2-n-heptyl-4-hydroxyquinoline N-oxide
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.12
-
2,3-dimethyl-1,4-naphthohydroquinone
-
-
0.0054
-
fumarate
P00363
succinate oxidation, pH 7.8, 30C
0.02
-
fumarate
-
wild-type, pH 7.0, 30C
0.03
-
fumarate
-
mutant E49A, pH 7.0, 30C
0.0018
-
menaquinone
-
mutant E29L, pH 7.0, 30C
0.004
-
menaquinone
-
wild-type, pH 7.0, 30C
0.0015
-
succinate
P00363
fumarate reduction, pH 7.8, 30C
0.11
-
succinate
-
mutant E49A, pH 7.0, 30C
0.22
-
succinate
-
mutant E49A, pH 7.0, 30C
0.55
-
succinate
-
wild-type, pH 7.0, 30C
0.64
-
succinate
-
pH 7.6, 30C
0.33
-
phenazine methosulfate
-
pH 7.6, 30C
additional information
-
succinate
P00363
fumarate reduction. 0.0013 mM, with ubiquinone, reaction of succinate-ubiquinone oxidase EC 1.3.5.1
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.3
-
fumarate
-
mutant K228R, cosubstrate menaquinone, pH 7.0, 30C
1
-
fumarate
-
mutant E49A, pH 7.0, 30C
2.3
-
fumarate
-
mutant E49F, cosubstrate menaquinone, pH 7.0, 30C
5
-
fumarate
-
mutant E49L, cosubstrate menaquinone, pH 7.0, 30C
20
-
fumarate
-
based on the content of the MW 79000 peptide, reduction of fumarate with dimethylnaphthohydroquinone
32
-
fumarate
-
mutant E49A, pH 7.0, 30C
133
-
fumarate
-
with 2,3-dimethyl-1,4-naphthohydroquinone, based on FAD content of enzyme
177
-
fumarate
P00363
succinate oxidation, pH 7.8, 30C
230
-
fumarate
-
wild-type, cosubstrate menaquinone, pH 7.0, 30C
250
-
fumarate
-
wild-type, pH 7.0, 30C
0.3
-
succinate
-
mutant E49L, cosubstrate menaquinone, pH 7.9, 30C
0.4
-
succinate
-
mutant E49F, cosubstrate menaquinone, pH 7.9, 30C
2.4
-
succinate
-
mutant E49A, pH 7.0, 30C
4
-
succinate
-
mutant E49A, pH 7.0, 30C
13
-
succinate
-
with 2,3-dimethyl-1,4-naphthoquinone, based on FAD content of enzyme
14
-
succinate
P00363
fumarate reduction, pH 7.8, 30C
15
-
succinate
-
wild-type, cosubstrate menaquinone, pH 7.9, 30C
30
-
succinate
-
wild-type, pH 7.0, 30C
80
-
succinate
-
reaction with phenazine methosulfate and 2,6-dichlorophenolindophenol
77
-
phenazine methosulfate
-
pH 7.6, 30C
additional information
-
succinate
P00363
fumarate reduction. 28 s-1, with ubiquinone, reaction of succinate-ubiquinone oxidase EC 1.3.5.1
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
97
-
fumarate
-
mutant E49A, pH 7.0, 30C
10622
232
-
fumarate
-
wild-type, pH 7.0, 30C
10622
11
-
succinate
-
mutant E49A, pH 7.0, 30C
16754
36
-
succinate
-
mutant E49A, pH 7.0, 30C
16754
54
-
succinate
-
wild-type, pH 7.0, 30C
16754
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.00006
-
2-n-heptyl-4-hydroxyquinoline-N-oxide
-
mutant E29L, pH 7.9, 30C
0.000075
-
2-n-heptyl-4-hydroxyquinoline-N-oxide
P00363
fumarate reduction, pH 7.8, 30C
0.0002
-
2-n-heptyl-4-hydroxyquinoline-N-oxide
-
wild-type, pH 7.0, 30C
0.0002
-
2-n-heptyl-4-hydroxyquinoline-N-oxide
P00363
succinate oxidation, pH 7.8, 30C
0.005
-
malonate
-
mutant E49A, pH 7.0, 30C
0.01
-
malonate
-
mutant E49A, pH 7.0, 30C
0.025
-
malonate
-
wild-type, pH 7.0, 30C
0.088
-
malonate
-
pH 7.6, 30C
0.00006
-
oxaloacetate
-
mutant E49A, pH 7.0, 30C
0.0001
-
oxaloacetate
-
mutant E49A, pH 7.0, 30C
0.0003
-
oxaloacetate
-
wild-type, pH 7.0, 30C
0.023
-
Pentachlorophenol
P00363
fumarate reduction, pH 7.8, 30C
0.037
-
Pentachlorophenol
P00363
succinate oxidation, pH 7.8, 30C
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.0076
-
-
electron acceptor 2,6-dichlorophenolindophenol, pH 7.6, 30C
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.8
-
P00363
assay at
7.8
-
-
with phenazine methosulfate-dichlorophenolindophenyl as electron acceptor
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
P00363
pH profile analysis, both EC 1.3.5.1 and 1.3.5.4 show a similar profile, suggesting that similar amino acid residues may be involved in quinol deprotonation and oxidation in Escherichia coli enzymes
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
30
-
P00363
assay at
30
-
-
oxidation of succinate, assay at; reduction of fumarate, assay at
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
the substrate and the dye-reactive sites of the enzyme face the inside of the cytoplasmic membrane
-
Manually annotated by BRENDA team
-
the SdhABE complex is membrane associated rather than tightly membrane bound
Manually annotated by BRENDA team
-
cytoplasmic membrane
Manually annotated by BRENDA team
P00363
membrane-bound, the catalytic domain is bound to the cytoplasmic membrane by 2 hydrophobic membrane anchor subunits that also form the sites of interaction with quinones
Manually annotated by BRENDA team
-
the formate-fumarate reductase electron transport system is localized in the membrane fraction of the bacterium, together with acid-extractable and covalently-bound FAD, menaquinone, iron-sulfur protein and h and c cytochromes
Manually annotated by BRENDA team
-
the hydrophilic subunits of the MFR complex are exported into the periplasm via the twin-arginine translocation (tat)-pathway
-
Manually annotated by BRENDA team
-
soluble fraction contains FMN, acid-extractable FAD, iron-sulfur protein and c cytochromes of the formate-fumarate reductase electron transport system
-
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Shewanella frigidimarina (strain NCIMB 400)
Shewanella oneidensis (strain MR-1)
Shewanella oneidensis (strain MR-1)
Shewanella oneidensis (strain MR-1)
Wolinella succinogenes (strain ATCC 29543 / DSM 1740 / LMG 7466 / NCTC 11488 / FDC 602W)
Wolinella succinogenes (strain ATCC 29543 / DSM 1740 / LMG 7466 / NCTC 11488 / FDC 602W)
Wolinella succinogenes (strain ATCC 29543 / DSM 1740 / LMG 7466 / NCTC 11488 / FDC 602W)
Wolinella succinogenes (strain ATCC 29543 / DSM 1740 / LMG 7466 / NCTC 11488 / FDC 602W)
Wolinella succinogenes (strain ATCC 29543 / DSM 1740 / LMG 7466 / NCTC 11488 / FDC 602W)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
110000
114000
-
excluding cytochrome b, calculation from FAD-content and subunit composition
160000
167000
-
including cytochrome b, calculation from FAD-content and subunit composition
200000
-
-
including cytochrome b, sedimentation equilibrium centrifugation
200000
-
-
calculated from the molar mass of the enzyme particle and its contents of Triton and phospholipid
260000
-
-
Blue-native PAGE
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
1 * 73234 + 1 * 28064 + 1 * 27097 Da calculated for flavoprotein, iron-sulfur protein, and cytochrome subunit. Complex is composed of three subunits, a 74 kDa flavoprotein that contains a covalently bound flavin adenine dinucleotide, a 28 kDa iron-sulfur cluster-containing polypeptide, and a 27 kDa transmembrane polypeptide, which is also the binding site of two b-type hemes and two menaquinones
oligomer
-
homodimeric complex of heterotrimers of A, B, and C subunits
tetramer
-
1 * 79000, FAD-binding subunit, 1 * 31000, Fe-S cluster containing subunit, 2 * 25000, cytochrome b containing subunits, SDS-PAGE
tetramer
-
fumarate reductase, membrane-extrinsic domain: 1 * 69000 + 1 * 27000, membrane-intrinsic domain: 1 * 15000 + 1 * 13000 containing cytochrome b, necessary for converting succinate dehydrogenase EC 1.3.99.1 into succinate-ubiquinone oxidoreductase
tetramer
P92507
QFR is composed of Fp, Ip, CybL and CybS subunits
trimer
-
enzyme containing cytochrome b, 1 * 79000 + 1 * 31000 + 1 * 25000
trimer
-
1 * 79000 + 1 * 31000 + 1 * 25000
trimer
-
homotrimeric complex of the heterotrimeric protomer
?
-
enzyme consists of a 65000 Da flavoprotein SdhA, a 29000 Da iron-sulfur protein SdhB, and a 19000 Da subunit SdhC containing two b-type cytochromes, SDS-PAGE
additional information
P00363
cf. EC 1.3.5.1, both complexes contain a catalytic domain, composed of a subunit with a covalently bound flavin cofactor, the dicarboxlyate binding site, and an iron-sulfur subunit, which contains three distince iron-sulfur clusters. The catalytic domain is bound to the cytoplasmic membrane by two hydrophobic membrane anchor subunits that also form the sites for interaction with quinones. The catalytic domain is highly conserved and reflect the biochemical and structural similarity of EC 1.3.5.1 (SQR) and 1.3.5.4 (QFR). SQR, in addition to differences in the type of quinones it uses as compared to QFR, is known to contain a single B556 heme moiety, showing to have bis-histidine axial ligation
additional information
P92507
the enzyme structure comprises four subunits and five co-factors, subunit structure comparisons, overview
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
purified native enzyme by dialysis method using a reservoir solution containing 15% w/v PEG 3350, 100 mM Tris-HCl, pH 8.4, 200 mM NaCl, 1 mM sodium malonate or fumarate, 0.06% w/v n-dodecyl ethylene glycol monoether C12E8, and 0.04% w/v n-dodecyl-bet-D-maltoside, 2-3 days, X-ray diffraction structure determination and analysis at 2.81-2.91 A resolution, molecular replacement
P92507
X-ray diffraction data up to 3.2 resolution
-
crystal structure of QFR to 3.3 A resolution. Enzyme contains two quinone species, presumably menaquinol, bound to the transmembrane-spanning region. The binding sites for the two quinone molecules are termed QP and QD, indicating their positions proximal, QP, or distal, QD, to the site of fumarate reduction in the hydrophilic flavoprotein and iron-sulfur protein subunits. Co-crystallization studies of the Escherichia coli QFR with the quinol-binding site inhibitors 2-heptyl-4-hydroxyquinoline-N-oxide and 2-[1-(p-chlorophenyl)ethyl] 4,6-dinitrophenol establish that both inhibitors block the binding of MQH2 at the QP site. In the structures with the inhibitor bound at QP, no density is observed at QD. The conserved acidic residue, Glu29 in subunit FrdC, in the Escherichia coli enzyme may act as a proton shuttle from the quinol during enzyme turnover
-
hanging drop vapor diffusion method, x-ray structure of mutant E49Q
-
PDB code: 1FUM, structure of the QFR monomer, with the covalently bound FAD cofactor, showing the iron-sulfur clusters [4Fe-4S], [3Fe-3S], and [2Fe-2S] and the two menaquinone molecules
P00363
the structure of the enzyme is determined at 2.2 A resolution by X-ray crystallography
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
native enzyme from muscle mitochondria by anion exchange chromatography
P92507
and reconstitution into proteoliposomes
-
using cell homogenate, Triton X-100 treatment and chromatography on hydroxyapatite and DEAE-Sepharose column
-
using solubilization with Triton X-100, chromatography on hydroxyapatite column and DEAE-Sephadex column, the enzyme elutes from the ion-exchange column in two forms, one containing and the other lacking cytochrome b
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
cloning of a mutant fumarate reductase operon
-
gene cluster frdABCD encoding 4 subunits, DNA and amino acid sequence analysis, overepression
P00363
expression in Escherichia coli
-
two structural genes coding for subunits of the enzyme cloned in Escherichia coli, polarity and organization of the genes
-
EXPRESSION
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
sdhABE operon is upregulated in an oxygen-limited environment as compared with microaerophilic laboratory conditions
-
the QFR complex provides electron transport during anaerobic cell growth conditions. The transcription of the frdABCD operon responds to environmental as well as internal cell signals to modulate gene expression. The transcription is coupled to that of the succinate-ubiquinone oxidase, EC 1.3.5.1, overview
P00363
fumarate reductase expression is repressed under conditions of growth during which electron transport to oxygen or to nitrate is possible
-
fumarate reductase is expressed under anaerobic growth conditions
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
C247
-
mutation in flavoprotein subunit FrdA. Increase in fumarate reduction rate, slight increase in succinate oxidation. Residue C247 of FrdA is responsible for the N-ethylmaleimide sensitivity shown by fumarate reductase but is not required for catalytic activity or the tight-binding of oxalacetate
E29F
-
mutation in subunit FrdC, dramatic decrease in enzymatic reactions with menaqunione. Elimination of the negative charge in E29 mutant enzymes results in significantly increased stabilization of both ubiquinone and menaquinone semiquinones
E29L
-
mutation in subunit FrdC, dramatic decrease in enzymatic reactions with menaqunione. Elimination of the negative charge in E29 mutant enzymes results in significantly increased stabilization of both ubiquinone and menaquinone semiquinones
E49A
-
decrease in catalytic efficiency of both fumarate reduction and succinate oxidation
E49Q
-
decrease in catalytic efficiency of both fumarate reduction and succinate oxidation
H232S
-
mutation in flavoprotein subunit FrdA. Decrease in fumarate reduction, strong decrease in succinate oxidation. Residue H232 is the general acid-base catalyst
H82R
-
menaquinone, ubiquinone and b-type cytochrome levels are present in normal amounts, the mutation alters the electron transfer properties of the iron-sulfur and flavin redox centers of the catalytic domain, functional electron flow from 2,3-dimethyl-1,4-naphthoquinone or from the electron transport chain is impaired, the mutant can be reduced normally by single-electron donors such as benzyl viologen
K228L
-
mutation in subunit FrdB. Residue K228 provides a strong hydrogen bond to menaquinone and is essential for reactions with both ubiquinone and menaquinone
R248H
-
mutation in flavoprotein subunit FrdA. Strong decrease both in fumarate reduction and in succinate oxidation
R248L
-
mutation in flavoprotein subunit FrdA. Strong decrease both in fumarate reduction and in succinate oxidation
A86H
-
in wild-type enzyme FAD is non-covalenly-bound. In the enzyme containing a mutant A86H flavoprotein subunit the FAD is covalently bound
E180Q
-
site-diirected mutagenesis, the mutant catalyzes the electron transfer from succinate to methylene blue, but not from 2,3-dimethyl-1,4-naphthoquinol to fumarate
E66Q
-
site-diirected mutagenesis, the mutant catalyzes the electron transfer from succinate to methylene blue, but not from 2,3-dimethyl-1,4-naphthoquinol to fumarate
H44E
-
site-diirected mutagenesis, although the H44E variant enzyme retains both heme groups, it is unable to catalyze quinol oxidation, the mutant catalyzes the electron transfer from succinate to methylene blue, with reduced activity compared to the wild-type enzyme but not from 2,3-dimethyl-1,4-naphthoquinol to fumarate
K228R
-
mutation in subunit FrdB. Residue K228 provides a strong hydrogen bond to menaquinone and is essential for reactions with both ubiquinone and menaquinone
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
additional information
P00363
His44 mutant contains non-covalently bound FAD and loose the ability to oxidize succinate
additional information
-
isolation of a mutant in the frdD gene encoding the hydrophic subunit of the fumarate reductase complex. In this mutant, fumarate reductase is not as tightly bound to the membrane. The mutation in the FrdD peptide causes an almost total loss of the ability of the enzyme to oxidize either menaquinol-6, or reduced benzyl viologen. The mutation does not impair the ability of the membrane-bound fumarate reductase complex to function with succinate as substrate
Renatured/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
menaquinone is an obligatory redox mediator of formate-fumarate reductase electron transport phosphorylation system. The activity is fully inhibited on the extraction of the menaquinone from the membrane fraction, and is reactivated on reincorporation of menaquinone into the membrane
-
the cleavage of the complex causes the complete loss of activity of fumarate reduction by dimethylnaphthohydroquinone, the activity can be restored by coprecipitation of the three subunits of MW: 79000 Da, 31000 Da and 25000 Da
-