1.16.3.4: cuproxidase
This is an abbreviated version!
For detailed information about cuproxidase, go to the full flat file.
Word Map on EC 1.16.3.4
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1.16.3.4
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multicopper
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laccase
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oxidases
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methionine-rich
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trinuclear
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2,6-dimethoxyphenol
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dioxygen
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bioelectrocatalysis
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ferroxidase
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cathodic
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ceruloplasmin
- 1.16.3.4
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multicopper
- laccase
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oxidases
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methionine-rich
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trinuclear
- 2,6-dimethoxyphenol
- dioxygen
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bioelectrocatalysis
- ferroxidase
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cathodic
- ceruloplasmin
Reaction
4 Cu+
+
4 H+
+
Synonyms
ceruloplasmin, copper efflux oxidase, Cu(I) oxidase, CueO, CuiD, cuprous oxidase, DA2_0547, fet3p, More, multicopper oxidase, multicopper oxidase CueO, YacK
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1.16.3.4 cuproxidaseAdvanced search results
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Engineering on EC 1.16.3.4 - cuproxidase
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PROTEIN VARIANTS 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
C500S
D439A
E506A
mutation results in the formation of a compensatory hydrogen bond network with one or two extra water molecules
E506I
mutation results in the complete shutdown of the hydrogen bond network leading to loss of enzymatic activities
E506Q
mutation results in the hydrogen bond network without the proton transport function
G304K
mutant shows markedly increased the laccase activity. Movements of the regulatory loop combined with the changes of the methionine-rich region may uncover the T1 Cu site allowing greater access of the substrate. Cuprous oxidase activity of mutant G304K is about 20% of wild-type activity, while the Km value is about 4times lower
M358S/M361S/M362S/M364S/M366S
mutation leads to an about 4fold reduction in kcat for Cu(I) oxidation
M358S/M361S/M362S/M368S/M376S
mutation leads to an about 4fold reduction in kcat for Cu(I) oxidation
M441L
mutation in labile copper coordination sphere. Mutation has affected copper incorporation into the T1 copper site
M510L
3.8-4.2 copper atoms per protein molecule, similar to wild-type
M510Q
3.8-4.2 copper atoms per protein molecule, similar to wild-type
P444A
P444A/D439A
mutation leads to a synergetic effect of the positive shift in the redox potential of the type I copper center and the increase in enzyme activity
P444A/M510Q
3.4 copper atoms per protein molecule, similar to wild-type
P444G
mutation results in positive shifts in the redox potential of this copper center and enhanced oxidase activity in CueO and in the region Pro357-His406 deletion mutant lacking a methionine-rich helical segment that covers the substrate-binding site
P444I
positive shift in the redox potential of this copper center and enhanced oxidase activity
P444L
positive shift in the redox potential of this copper center and enhanced oxidase activity
M358S/M361S/M362S/M364S/M366S
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mutation leads to an about 4fold reduction in kcat for Cu(I) oxidation
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M455L
mutation in T1 Cu site, incorporation of 3-4 copper atoms, similar to wild-type. Mutation results in an increase of 100 mV in the O2 reduction potential, while the enzymatic activity for ABTS oxidation is decreased
M456A
mutation in T1 Cu site, incorporation of 3-4 copper atoms, similar to wild-type
additional information
C500S
mutant designed to incapacitate binding of the T1 copper, mutant protein does not support resistance to copper toxicity when placed in a cell line missing a functional CueO gene
D439A
3.8-4.2 copper atoms per protein molecule, similar to wild-type
D439A
mutation in labile copper coordination sphere, 3-4fold increase in Km value for Cu2+
D439A
positive shift in the redox potential of this copper center and enhanced oxidase activity
P444A
3.8-4.2 copper atoms per protein molecule, similar to wild-type
P444A
positive shift in the redox potential of this copper center and enhanced oxidase activity
additional information
deletion of the region Pro357-His406 comprising a methionine-rich helical segment that covers the substrate-binding site and replacement with a Gly-Gly linker. The scaffold of the CueO molecule and metal-binding sites are reserved in the mutant. The high thermostability of the protein molecule and its spectroscopic and magnetic properties are also conserved after truncation. The cuprous oxidase activity of the mutant is reduced to about 10% that of recombinant CueO due to the decrease in the affinity of the labile Cu site for Cu(I) ions
additional information
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deletion of the region Pro357-His406 comprising a methionine-rich helical segment that covers the substrate-binding site and replacement with a Gly-Gly linker. The scaffold of the CueO molecule and metal-binding sites are reserved in the mutant. The high thermostability of the protein molecule and its spectroscopic and magnetic properties are also conserved after truncation. The cuprous oxidase activity of the mutant is reduced to about 10% that of recombinant CueO due to the decrease in the affinity of the labile Cu site for Cu(I) ions
additional information
mutations at Pro444 to construct a second NH-S hydrogen bond between the backbone amide and coordinating Cys500 thiolate of the type I copper result in positive shifts in the redox potential of this copper center and enhanced oxidase activity in CueO. Pro444 mutations limit the incorporation of copper ions into the trinuclear copper center. The activities of both CueO and the region Pro357-His406 deletion mutant are also enhanced by mutations to break down the hydrogen bond between the imidazole group of His443 that is coordinated to the type I copper and the beta-carboxy group of Asp439 that is located in the outer sphere of the type I copper center. The characteristics of the Cu(II)-S(Cys) bond are only minimally perturbed by mutations involving formation or disruption of a hydrogen bond from the coordinating groups to the type I copper
additional information
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mutations at Pro444 to construct a second NH-S hydrogen bond between the backbone amide and coordinating Cys500 thiolate of the type I copper result in positive shifts in the redox potential of this copper center and enhanced oxidase activity in CueO. Pro444 mutations limit the incorporation of copper ions into the trinuclear copper center. The activities of both CueO and the region Pro357-His406 deletion mutant are also enhanced by mutations to break down the hydrogen bond between the imidazole group of His443 that is coordinated to the type I copper and the beta-carboxy group of Asp439 that is located in the outer sphere of the type I copper center. The characteristics of the Cu(II)-S(Cys) bond are only minimally perturbed by mutations involving formation or disruption of a hydrogen bond from the coordinating groups to the type I copper
additional information
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deletion of the longest methionine-rich surface loop lowers bound extra Cu(I) from 9 in the wild-type enzyme to 2-3 Cu(I) in deletion mutants
additional information
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deletion of the longest methionine-rich surface loop lowers bound extra Cu(I) from 9 in the wild-type enzyme to 2-3 Cu(I) in deletion mutants
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additional information
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truncated enzyme with a deleted methionine-rich C-terminal tail region shows a decreased turnover and a slightly lower Km value. Ki (Cu+) value increased compared to wild-type. Catalytic efficacy similar to wild-type
