Information on EC 1.10.3.10 - ubiquinol oxidase (H+-transporting)

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The expected taxonomic range for this enzyme is: Proteobacteria

EC NUMBER
COMMENTARY
1.10.3.10
-
RECOMMENDED NAME
GeneOntology No.
ubiquinol oxidase (H+-transporting)
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
2 ubiquinol + O2 + n H+[side 1] = 2 ubiquinone + 2 H2O + n H+[side 2]
show the reaction diagram
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
D-lactate to cytochrome bo oxidase electron transport
-
-
glycerol-3-phosphate to cytochrome bo oxidase electron transfer
-
-
NADH to cytochrome bo oxidase electron transfer I
-
-
NADH to cytochrome bo oxidase electron transfer II
-
-
proline to cytochrome bo oxidase electron transfer
-
-
pyruvate to cytochrome bo oxidase electron transfer
-
-
succinate to cytochrome bo oxidase electron transfer
-
-
oxidative phosphorylation
-
-
SYSTEMATIC NAME
IUBMB Comments
ubiquinol:O2 oxidoreductase (H+-transporting)
Contains a dinuclear centre comprising two hemes, or heme and copper. This terminal oxidase enzyme generates proton motive force by two mechanisms: (1) transmembrane charge separation resulting from utilizing protons and electrons originating from opposite sides of the membrane to generate water, and (2) active pumping of protons across the membrane. The bioenergetic efficiency (the number of charges driven across the membrane per electron used to reduce oxygen to water) depends on the enzyme; for example, for the bo3 oxidase it is 2, while for the bd-II oxidase it is 1. cf. EC 1.10.3.14, ubiquinol oxidase (electrogenic, non H+-transporting).
SYNONYMS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
bd-type quinol oxidase
-
-
bd-type quinol oxidase
Escherichia coli GO105
-
-
-
bo-type ubiquinol oxidase
-
-
bo-type ubiquinol oxidase
Escherichia coli GO103/pMFO2, Escherichia coli RG145, Escherichia coli ST4676
-
-
-
Cyt-bo3
-
-
cytochrome bd-I oxidase
-
-
-
-
cytochrome bd-I quinol oxidase
-
-
cytochrome bd-I quinol oxidase
-
-
-
cytochrome bd-II oxidase
-
-
-
-
cytochrome bd-II quinol oxidase
-
-
cytochrome bd-II quinol oxidase
-
-
-
cytochrome bo
Escherichia coli GO103/pHN3795-1, Escherichia coli GO103/pMFO2, Escherichia coli RG145, Escherichia coli ST4676
-
-
-
cytochrome bo oxidase
-
-
-
-
cytochrome bo quinol oxidase
-
-
cytochrome bo quinol oxidase
Escherichia coli RG145
-
-
-
cytochrome bo terminal oxidase
-
-
cytochrome bo terminal oxidase
Escherichia coli RG145
-
-
-
cytochrome bo ubiquinol oxidase
-
-
cytochrome bo ubiquinol oxidase
Escherichia coli GL101
-
;
-
cytochrome bo ubiquinol oxidase
Escherichia coli GLlOl, Escherichia coli JM103
-
-
-
cytochrome bo ubiquinol oxidase
-
-
cytochrome bo-type ubiquinol oxidase
-
-
cytochrome bo-type ubiquinol oxidase
Escherichia coli GO103
-
;
-
cytochrome bo-type ubiquinol oxidase
-
-
cytochrome bo3
-
-
cytochrome bo3
-
cytochrome bo3 oxidase
-
-
cytochrome bo3 ubiquinol oxidase
-
-
cytochrome bo3 ubiquinol oxidase
Escherichia coli C43 (DE3), Escherichia coli GO105
-
-
-
cytochrome o complex
-
-
cytochrome o complex
Escherichia coli RG145
-
-
-
QObo3
Escherichia coli C43 (DE3)
-
-
-
ubiquinol oxidase
-
ubiquinol oxidase:O2 reductase
-
-
ubiquinol oxidase:O2 reductase
Escherichia coli RG145
-
-
-
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Escherichia coli C43 (DE3)
-
-
-
Manually annotated by BRENDA team
Escherichia coli GL101
-
-
-
Manually annotated by BRENDA team
Escherichia coli GLlOl
-
-
-
Manually annotated by BRENDA team
Escherichia coli GO103
-
-
-
Manually annotated by BRENDA team
Escherichia coli GO103/pHN3795-1
-
-
-
Manually annotated by BRENDA team
Escherichia coli GO103/pMFO2
-
-
-
Manually annotated by BRENDA team
Escherichia coli GO105
-
-
-
Manually annotated by BRENDA team
Escherichia coli JM103
-
-
-
Manually annotated by BRENDA team
Escherichia coli RG145
-
-
-
Manually annotated by BRENDA team
Escherichia coli ST4676
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
malfunction
-
loss of cytochrome bd-I oxidase subunit II (gene cydB) causes diminished respiration rates, impaired motility and enhanced acid resistance. CydB cells contain elevated heme d, particularly at low pH. The GABA/glutamate gadC antiporter is highly up-regulated in cydB cells. Eschrichia coli can compensate for the loss of cytochrome bd-I activity
malfunction
-
deletion of the cydAB genes for cytochrome bd has no obvious influence on growth, whereas the lack of the cyoBACD genes for cytochrome bo3 severely reduced the growth rate and the cell yield
malfunction
-
loss of cytochrome bd-I oxidase subunit II (gene cydB) causes diminished respiration rates, impaired motility and enhanced acid resistance. CydB cells contain elevated heme d, particularly at low pH. The GABA/glutamate gadC antiporter is highly up-regulated in cydB cells. Eschrichia coli can compensate for the loss of cytochrome bd-I activity
-
metabolism
-
Gluconobacter oxydans oxidizes a variety of substrates in the periplasm by membrane-bound dehydrogenases, which transfer the reducing equivalents to ubiquinone. Two quinol oxidases, cytochrome bo3 and cytochrome bd, then catalyze transfer of the electrons from ubiquinol to molecular oxygen
physiological function
-
the ubiquinol oxidase, cytochrome b03, of Escherichia coli is a member of the respiratory heme-copper oxidase family and conserves energy from the reduction of dioxygen to water by translocation of protons across the bacterial membrane
physiological function
-
cytochrome bo3 ubiquinol oxidase serves as part of a proton loading site that regulates proton translocation across the protein matrix of the enzyme
physiological function
-
the bo-type ubiquinol oxidase is functioning as a proton pump
physiological function
-
cytochrome bo-type ubiquinol oxidase in the aerobic respiratory chain of Escherichia coli catalyzes the reduction of dioxygen to water with ubiquinol-8, and utilizes the redox reactions to drive vectorial translocation of protons across the cytoplasmic membrane
physiological function
-
cytochrome bo is a four-subunit quinol oxidase in the aerobic respiratory chain of Escherichia coli and functions as a redox-coupled proton pump
physiological function
-
cytochrome bd-II-mediated quinol oxidation prevents the accumulation of NADH, whereas GABA synthesis/antiport maintains the proton motive force for ATP production
physiological function
cytochrome bo3 ubiquinol oxidase from Escherichia coli is a four-subunit heme-copper oxidase that catalyzes the four-electron reduction of O2 to water and functions as a proton pump
physiological function
-
cytochrome bo3 might be a rate-limiting factor of the respiratory chain
physiological function
-
cytochrome bd-II-mediated quinol oxidation prevents the accumulation of NADH, whereas GABA synthesis/antiport maintains the proton motive force for ATP production
-
physiological function
Escherichia coli GO103
-
cytochrome bo-type ubiquinol oxidase in the aerobic respiratory chain of Escherichia coli catalyzes the reduction of dioxygen to water with ubiquinol-8, and utilizes the redox reactions to drive vectorial translocation of protons across the cytoplasmic membrane
-
physiological function
Escherichia coli GO103/pMFO2, Escherichia coli ST4676
-
cytochrome bo is a four-subunit quinol oxidase in the aerobic respiratory chain of Escherichia coli and functions as a redox-coupled proton pump
-
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
decylubiquinol + O2 + H+/in
decylubiquinone + H2O + H+/out
show the reaction diagram
-
-
-
?
N,N,N',N'-tetramethylphenylene diamine + O2
?
show the reaction diagram
-
-
-
-
?
ubiquinol + O2 + H+/in
ubiquinone + H2O + H+/out
show the reaction diagram
-
-
-
?
ubiquinol + O2 + H+/in
ubiquinone + H2O + H+/out
show the reaction diagram
Escherichia coli, Escherichia coli GO103/pMFO2
-
-
-
?
ubiquinol-1 + O2
ubiquinone-1 + H2O
show the reaction diagram
-
-
-
?
ubiquinol-1 + O2 + H+/in
ubiquinone-1 + H2O + H+/out
show the reaction diagram
-
-
-
?
ubiquinol-1 + O2 + H+/in
ubiquinone-1 + H2O + H+/out
show the reaction diagram
-
-
-
?
ubiquinol-1 + O2 + H+/in
ubiquinone-1 + H2O + H+/out
show the reaction diagram
-
-
-
?
ubiquinol-1 + O2 + H+/in
ubiquinone-1 + H2O + H+/out
show the reaction diagram
-
-
-
?
ubiquinol-1 + O2 + H+/in
ubiquinone-1 + H2O + H+/out
show the reaction diagram
-
-
-
?
ubiquinol-1 + O2 + H+/in
ubiquinone-1 + H2O + H+/out
show the reaction diagram
-
-
-
?
ubiquinol-1 + O2 + H+/in
ubiquinone-1 + H2O + H+/out
show the reaction diagram
-
-
-
?
ubiquinol-1 + O2 + H+/in
ubiquinone-1 + H2O + H+/out
show the reaction diagram
-
-
-
?
ubiquinol-1 + O2 + H+/in
ubiquinone-1 + H2O + H+/out
show the reaction diagram
-
-
-
?
ubiquinol-1 + O2 + H+/in
ubiquinone-1 + H2O + H+/out
show the reaction diagram
-
-
?
ubiquinol-1 + O2 + H+/in
ubiquinone-1 + H2O + H+/out
show the reaction diagram
Escherichia coli GL101
-
-
-
?
ubiquinol-1 + O2 + H+/in
ubiquinone-1 + H2O + H+/out
show the reaction diagram
Escherichia coli GL101
-
-
-
?
ubiquinol-1 + O2 + H+/in
ubiquinone-1 + H2O + H+/out
show the reaction diagram
Escherichia coli GO103
-
-
-
?
ubiquinol-1 + O2 + H+/in
ubiquinone-1 + H2O + H+/out
show the reaction diagram
Escherichia coli GO103/pHN3795-1
-
-
-
?
ubiquinol-1 + O2 + H+/in
ubiquinone-1 + H2O + H+/out
show the reaction diagram
Escherichia coli ST4676
-
-
-
?
ubiquinol-1 + O2 + H+/in
ubiquinone-1 + H2O + H+/out
show the reaction diagram
Escherichia coli C43 (DE3)
-
-
-
?
ubiquinol-1 + O2 + H+/in
ubiquinone-1 + H2O + H+/out
show the reaction diagram
Escherichia coli RG145
-
-
-
?
ubiquinol-1 + O2 + H+/in
ubiquinone-1 + H2O + H+/out
show the reaction diagram
Escherichia coli RG145
-
-
-
?
ubiquinol-8 + O2 + H+/in
ubiquinone-8 + H2O + H+/out
show the reaction diagram
Escherichia coli, Escherichia coli GO103
-
-
-
?
duroquinol + O2 + H+/in
duroquinone + H2O + H+/out
show the reaction diagram
-
-
-
?
additional information
?
-
-
a significant interaction is observed only between Vitreoscilla hemoglobin and subunit I of cytochrome bo
-
-
-
additional information
?
-
-
electron transfer between hemes d and b595 is not electrogenic, although heme b595 is the major electron acceptor for heme d during the backflow, and therefore is not likely to be accompanied by net H+ uptake or release
-
-
-
additional information
?
-
-
horse heart reduced cytochrome c is not oxidized by this enzyme
-
-
-
additional information
?
-
-
the enzyme does not interact with cytochrome c
-
-
-
additional information
?
-
-
enzymatic activity of Cyt-bo3 was estimated by measuring consumption of oxygen for ubiquinol oxidation
-
-
-
additional information
?
-
Escherichia coli JM103
-
a significant interaction is observed only between Vitreoscilla hemoglobin and subunit I of cytochrome bo
-
-
-
additional information
?
-
Escherichia coli GO105
-
electron transfer between hemes d and b595 is not electrogenic, although heme b595 is the major electron acceptor for heme d during the backflow, and therefore is not likely to be accompanied by net H+ uptake or release
-
-
-
additional information
?
-
Escherichia coli RG145
-
the enzyme does not interact with cytochrome c
-
-
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
cytochrome
-
the oxidase contains cytochrome b, cytochrome o
-
heme
-
the enzyme possesses two hemes (18.1 ng/mg)
heme
-
heme b562
heme
-
the high-spin heme is magnetically coupled to a copper, CuB, forming a binuclear center which is the site of oxygen reduction to water
heme
-
the ubiquinol oxidase cytochrome b03 of Escherichia coli is a member of the respiratory heme-copper oxidase family
heme
-
the purified oxidase contains both protoheme and heme O
heme
-
the enzyme contains 2 mol of heme, one or both of which are heme o
heme
-
the enzyme contains a low-spin b-like heme and a high-spin 6-like heme, designated cytochromes b and o respectively
heme
-
the enzyme contains a hexa-coordinated low-spin heme, heme B and a penta-coordinated high-spin heme, heme O
heme
-
hemes B and O
heme
-
low-spin heme b and high-spin heme o in subunit I
heme
-
low-spin heme b and high-spin heme o
heme
-
heme d and heme b558
ubiquinol-8
-
-
ubiquinone-8
-
the enzyme contains one equivalent of ubiquinone-8
ubiquinone-8
-
the bound ubiquinone at the QH site of cytochrome bo is essential for the catalytic turnover of the oxidase reactions, but it is not necessary for re-reduction of ferric heme b after the heme b-to-heme o electron transfer under flow-flash conditions
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
copper
-
the high-spin heme is magnetically coupled to a copper, CuB, forming a binuclear center which is the site of oxygen reduction to water
copper
-
the enzyme has a CuB site
copper
-
the enzyme contains a copper ion in subunit I
copper
-
the enzyme is a heme-copper terminal oxidase
copper
-
the enzyme contains CuB
copper
the enzyme is a four-subunit heme-copper oxidase containing CuB
Cu
-
copper-containing enzyme
Cu2+
-
the pure four-subunit enzyme contains one equivalent of copper (7.5 ng/mg)
Cu2+
-
the ubiquinol oxidase cytochrome b03 of Escherichia coli is a member of the respiratory heme-copper oxidase family
Cu2+
-
the enzyme contains 6.78 nmol copper per mg of protein
Cu2+
-
the oxidase contains copper atoms
Fe
-
the pure four-subunit enzyme contains two equivalents of iron (19.5 ng/mg)
additional information
-
the enzyme contains no zinc. The equivalent of CuA of the aa3-type cytochrome c oxidases is absent in this quinol oxidase
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
2-heptyl-4-hydroxyquinoline N-oxide
-
-
2-n-heptyl-4-hydroxyquinoline N-oxide
-
-
aurachin C1-10
-
competitive inhibitor
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
diphosphatidyl glycerol
-
activates the enzyme with the optimum concentration at 0.04 mg/ml
n-dodecyl octaethyleneglycol monoether
-
effective activator
N-lauroylsarcosine
-
effective activator
Zwittergent 3-14
-
effective activator
liponox DCH
-
the activity is saturated at 0.01-0.02% (v/v) Liponox DCH
-
additional information
-
the enzyme is not activated by sodium cholate, phosphatidyl glycerol, phosphatidyl ethanolamine and soybean phospholipids
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.018
ubiquinol-1
wild type enzyme, when Triton X-100 is used as detergent in the isolation of the enzyme, in 50 mM Tris-HCl, pH 7.4, at 25C
0.023
ubiquinol-1
mutant enzyme Q101N, when Triton X-100 is used as detergent in the isolation of the enzyme, in 50 mM Tris-HCl, pH 7.4, at 25C
0.024
ubiquinol-1
mutant enzyme Q101N, when 0.02% (v/v) n-dodecyl beta-D-maltoside is used as detergent in the isolation of the enzyme, in 50 mM Tris-HCl, pH 7.4, at 25C
0.045
ubiquinol-1
wild type enzyme, when 0.02% (v/v) n-dodecyl beta-D-maltoside is used as detergent in the isolation of the enzyme, in 50 mM Tris-HCl, pH 7.4, at 25C
0.052
ubiquinol-1
-
mutant enzyme D188N, at 37C, pH 7.0
0.053
ubiquinol-1
-
wild type enzyme, at 37C, pH 7.0
0.056
ubiquinol-1
-
mutant enzyme D188A, at 37C, pH 7.0
0.06
ubiquinol-1
-
mutant enzyme R257Q, at 37C, pH 7.0
0.175
ubiquinol-1
mutant enzyme H98N, when 0.02% (v/v) n-dodecyl beta-D-maltoside is used as detergent in the isolation of the enzyme, in 50 mM Tris-HCl, pH 7.4, at 25C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
300
ubiquinol-1
Escherichia coli
-
in 60 mM Tris-HCl (pH 7.5), at 25C
341
ubiquinol-1
Escherichia coli
-
in 60 mM Tris-HCl (pH 7.5), at 25C
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.00074
2-heptyl-4-hydroxyquinoline N-oxide
-
wild type enzyme, at 37C, pH 7.0
0.000012
aurachin C1-10
-
mutant enzyme R257Q, at 37C, pH 7.0
0.000015
aurachin C1-10
-
wild type enzyme, at 37C, pH 7.0
0.000025
aurachin C1-10
-
mutant enzyme D188A, at 37C, pH 7.0
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0032
2-n-heptyl-4-hydroxyquinoline N-oxide
Vibrio alginolyticus
-
n 25 mM potassium phosphate (pH 7.5), at 30C
0.0064
KCN
Vibrio alginolyticus
-
in 25 mM potassium phosphate (pH 7.5), at 30C
0.0034
ZnSO4
Vibrio alginolyticus
-
in 25 mM potassium phosphate (pH 7.5), at 30C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
960
-
at 37C, pH not specified in the publication
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
Escherichia coli GO103, Escherichia coli RG145, Escherichia coli ST4676
-
-
-
-
Manually annotated by BRENDA team
Escherichia coli GL101
-
;
-
Manually annotated by BRENDA team
Escherichia coli JM103, Escherichia coli RG145
-
-
-
Manually annotated by BRENDA team
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dimer
-
x-ray crystallography
dimer
Escherichia coli GO105
-
x-ray crystallography
-
heterodimer
-
1 * 79000 + 1 * 36000 + 1 * 13000, SDS-PAGE
heterotetramer
-
1 * 58000 + 1 * 33000 + 1 * 22000 + 1 * 17000, SDS-PAGE
heterotetramer
Escherichia coli RG145
-
1 * 58000 + 1 * 33000 + 1 * 22000 + 1 * 17000, SDS-PAGE
-
tetramer
-
-
tetramer
x-ray crystallography
tetramer
Escherichia coli GO103/pMFO2
-
-
-
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging drop vapor diffusion method, using 9-10% (w/v) PEG 1500, 100 mM HEPES pH 7.0, 100 mM NaCl, 100 mM MgCl2, 5% (v/v) ethanol
-
hanging drop vapor diffusion method, using 9-10% (w/v) PEG 1500, 100 mM NaCl, 100 mM MgCl2 and 5% (v/v) ethanol
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DEAE-Sepharose column chromatography
-
Ni-NTA column chromatography
-
Ni2+-NTA column chromatography and MonoQ column chromatography
-
Ni2+-NTA-agarose column chromatography, gel filtration
-
recombinant C-terminally His6-tagged on subunit II Cyt-bo3 from Escherichia coli strain C43 by nickel affinity chromatography
-
DEAE-Sephacel gel filtration, DEAE-5PW gel filtration, and Sephacryl S-300 gel filtration
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli RG 129 cells
-
expressed in Escherichia coli RG129 cells
-
recombinant expression of Cyt-bo3 samples in Escherichia coli strain C43, each plasmid encodes the cyoABCDE operon expressing Cyt-bo3 with a 6-His tag on the C-terminus of subunit II. Functional cell-free in vitro expression of Cyt-bo3, employing T7 promoter, and insertion into the artificial membrane
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
D135E
-
the mutant shows 45% activity compared to the wild type enzyme
D135K
-
the mutant is deficient in proton pumping (23% activity compared to the wild type enzyme)
D135N
-
the mutant shows 45% activity compared to the wild type enzyme, with proton pumping decoupled from the electron-transfer activity
D135N
-
the mutant is deficient in proton pumping (45% activity compared to the wild type enzyme)
D135N
-
the mutations specifically eliminates the CuB center from the oxidase complex
D188N
-
the mutant shows 53% activity compared to the wild type enzyme
D188N
-
the mutant possesses 100% copper and 81% cytochrome o compared to the wild type enzyme
D256N
-
the mutant shows 25% activity compared to the wild type enzyme
D256N
-
the mutant possesses 103% copper and 74% cytochrome o compared to the wild type enzyme
D407N
-
the mutant shows 31% activity compared to the wild type enzyme
D407N
-
the mutation affects the CO-binding by the heme-copper binuclear center
D75E
-
135% activity compared to the wild type enzyme
D75H
-
4% activity compared to the wild type enzyme
D75N
-
inactive
D75N
the mutation inhibits activity by 99%
D75R
-
inactive
E286A
-
inactive
E286C
-
the mutant shows 3% activity compared to the wild type enzyme as a result of the inhibition of proton transfer from the D-channel
E286D
-
the mutant retains 31% of the wild type activity
E286D
-
the mutant does not show significant perturbations on the redox metal centers even though it is still inactive
E286Q
-
the mutant shows 69% activity compared to the wild type enzyme
E286Q
-
the mutant shows 4% activity compared to the wild type enzyme and is unable to bind azide ions
E286Q
-
the mutations specifically eliminates the CuB center from the oxidase complex
E445A
-
heme b595 is present in the E445A mutant. Formation of the oxoferryl state in the mutant is about 100fold slower than in the wild type enzyme. The E445A substitution does not affect intraprotein electron re-equilibration after the photolysis of CO bound to ferrous heme d in the one-electron-reduced enzyme. The mutation does not affect membrane potential generation coupled to intramolecular electron redistribution between hemes d2+ and b558
E540Q
-
the mutation affects the CO-binding by the heme-copper binuclear center
F112L
-
the mutation does not affect the in vivo activity
F113L
-
the mutation does not affect the in vivo activity
F138G
-
the mutant shows 63% proton-translocating activity compared to the wild type enzyme
F138R
-
the mutant shows 55% proton-translocating activity compared to the wild type enzyme
F208L
-
the mutation does not affect the in vivo activity
F295L
-
the mutation does not affect the in vivo activity
F336L
-
the mutation does not affect the in vivo activity
F347L
-
the mutation does not affect the in vivo activity
F348L
-
4.8% activity compared to the wild type enzyme
F391L
-
the mutation does not affect the in vivo activity
F415W
-
the mutation does not affect the in vivo activity
F420L
-
the mutation does not affect the in vivo activity
G132A
-
the mutant shows wild type proton-translocation activity (113% activity compared to the wild type enzyme)
G132D/D135N
-
the mutant shows 66% proton-translocating activity compared to the wild type enzyme
G132R
-
the mutant shows wild type proton-translocation activity
H333C
-
nonfunctional enzyme
H333L
-
the mutation eliminates the magnetic coupling between heme o and CuB leading to a nonfunctional enzyme
H333N
-
nonfunctional enzyme
H333Q
-
nonfunctional enzyme
H334L
-
the mutation eliminates the magnetic coupling between heme o and CuB leading to a nonfunctional enzyme
H334M
-
nonfunctional enzyme
H98N
-
1% activity compared to the wild type enzyme
H98N
the mutation inhibits activity by 97%
H98S
-
2% activity compared to the wild type enzyme
H98T
-
1% activity compared to the wild type enzyme
K362D/Dl35K
-
the mutant is devoid of redox activity
K362L
-
catalytically inactive
K362M
-
catalytically inactive
K362Q
-
catalytically inactive
K362Q
-
the mutation affects the CO-binding by the heme-copper binuclear center
K55Q
-
the mutant possesses 100% copper and 73% cytochrome o compared to the wild type enzyme
M353A
-
the mutant shows substantial activity
N124D
-
the mutant is deficient in proton pumping (56% activity compared to the wild type enzyme)
N124D/D135N
-
the mutant shows 21% proton-translocating activity compared to the wild type enzyme
N124H
-
the mutant is deficient in proton pumping (16% activity compared to the wild type enzyme)
N142D
-
the mutant is deficient in proton pumping (48% activity compared to the wild type enzyme)
N142D/D135N
-
the mutant shows 33% proton-translocating activity compared to the wild type enzyme
N142Q
-
the mutant shows wild type proton-translocation activity (109% activity compared to the wild type enzyme)
N142V
-
the mutant is deficient in proton pumping (22% activity compared to the wild type enzyme)
P128A
-
the mutant shows wild type proton-translocation activity (115% activity compared to the wild type enzyme)
P128D/D135N
-
inactive
P139E/D135N
-
the mutant shows 95% proton-translocating activity compared to the wild type enzyme
P358A
-
the mutant shows substantial activity
Pl39A
-
the mutant shows wild type proton-translocation activity (67% activity compared to the wild type enzyme)
Pl39E
-
the mutant shows wild type proton-translocation activity (46% activity compared to the wild type enzyme)
Q101N
-
5% activity compared to the wild type enzyme
Q101N
the mutation inhibits activity by 75% and causes a 10fold increase in the apparent KM for ubiquinol-1
R134P
-
the mutant shows 112% proton-translocating activity compared to the wild type enzyme
R257Q
-
the mutations specifically eliminates the CuB center from the oxidase complex
R481L
-
nonfunctional mutant
R481Q
-
the mutant is fully functional
R481Q
-
the mutant possesses 91% copper and 73% cytochrome o compared to the wild type enzyme
R482Q
-
the mutant possesses 82% copper and 100% cytochrome o compared to the wild type enzyme
R71D
-
inactive
R71D/D75R
-
inactive
R71K
-
inactive
R71L
the mutation inhibits activity by 99%
R71Q
-
inactive
R71Q
the mutation inhibits activity by 99%
R80Q
-
the mutation causes loss of a diagnostic peak for low-spin heme b in the 77 K redox difference spectrum
T352A
-
catalytically inactive
T352N
-
the mutant shows substantial activity
T352S
-
the mutant shows substantial activity
T359A
-
catalytically inactive
T359S
-
the mutant shows almost wild type activity
W147L
-
the mutation does not affect the in vivo activity
W280L
-
67% activity compared to the wild type enzyme
W282F
-
the mutation does not affect the in vivo activity
W331L
-
19% activity compared to the wild type enzyme
Y173F
-
the mutant possesses 91% copper and 108% cytochrome o compared to the wild type enzyme
Y288F
-
the mutations specifically eliminates the CuB center from the oxidase complex
Y288L
-
0.3% activity compared to the wild type enzyme
Y61F
-
the mutation does not affect the in vivo activity
E286C
Escherichia coli C43 (DE3)
-
the mutant shows 3% activity compared to the wild type enzyme as a result of the inhibition of proton transfer from the D-channel
-
D135N
Escherichia coli GL101
-
the mutant shows 45% activity compared to the wild type enzyme, with proton pumping decoupled from the electron-transfer activity
-
D256N
Escherichia coli GL101
-
the mutant shows 25% activity compared to the wild type enzyme
-
D407N
Escherichia coli GL101
-
the mutant shows 31% activity compared to the wild type enzyme
-
E286A
Escherichia coli GL101
-
inactive
-
E286Q
Escherichia coli GL101
-
the mutant shows 69% activity compared to the wild type enzyme
-
K362M
Escherichia coli GL101
-
catalytically inactive
-
K362Q
Escherichia coli GL101
-
catalytically inactive
-
T352A
Escherichia coli GL101
-
catalytically inactive
-
T352N
Escherichia coli GL101
-
the mutant shows substantial activity
-
T359A
Escherichia coli GL101
-
catalytically inactive
-
H333C
Escherichia coli GLlOl
-
nonfunctional enzyme
-
H333L
Escherichia coli GLlOl
-
the mutation eliminates the magnetic coupling between heme o and CuB leading to a nonfunctional enzyme
-
H333N
Escherichia coli GLlOl
-
nonfunctional enzyme
-
H333Q
Escherichia coli GLlOl
-
nonfunctional enzyme
-
H334L
Escherichia coli GLlOl
-
the mutation eliminates the magnetic coupling between heme o and CuB leading to a nonfunctional enzyme
-
E286A
Escherichia coli GO103
-
inactive
-
E286D
Escherichia coli GO103
-
the mutant retains 31% of the wild type activity
-
E286Q
Escherichia coli GO103
-
the mutant shows 4% activity compared to the wild type enzyme and is unable to bind azide ions
-
F113L
Escherichia coli GO103/pMFO2
-
the mutation does not affect the in vivo activity
-
F295L
Escherichia coli GO103/pMFO2
-
the mutation does not affect the in vivo activity
-
F347L
Escherichia coli GO103/pMFO2
-
the mutation does not affect the in vivo activity
-
F420L
Escherichia coli GO103/pMFO2
-
the mutation does not affect the in vivo activity
-
Y61F
Escherichia coli GO103/pMFO2
-
the mutation does not affect the in vivo activity
-
E445A
Escherichia coli GO105
-
heme b595 is present in the E445A mutant. Formation of the oxoferryl state in the mutant is about 100fold slower than in the wild type enzyme. The E445A substitution does not affect intraprotein electron re-equilibration after the photolysis of CO bound to ferrous heme d in the one-electron-reduced enzyme. The mutation does not affect membrane potential generation coupled to intramolecular electron redistribution between hemes d2+ and b558
-
D407N
Escherichia coli ST4676
-
the mutation affects the CO-binding by the heme-copper binuclear center
-
E286Q
Escherichia coli ST4676
-
the mutations specifically eliminates the CuB center from the oxidase complex
-
K55Q
Escherichia coli ST4676
-
the mutant possesses 100% copper and 73% cytochrome o compared to the wild type enzyme
-
R80Q
Escherichia coli ST4676
-
the mutation causes loss of a diagnostic peak for low-spin heme b in the 77 K redox difference spectrum
-
Y173F
Escherichia coli ST4676
-
the mutant possesses 91% copper and 108% cytochrome o compared to the wild type enzyme
-
additional information
-
plasmid-based overexpression of cyoBACD leads to increased growth rates and growth yields, both in the wild-type and the DELTAcyoBACD mutant, suggesting that cytochrome bo3 might be a rate-limiting factor of the respiratory chain