Information on EC 1.16.3.1 - ferroxidase and Organism(s) Homo sapiens

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


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EC NUMBER
COMMENTARY hide
1.16.3.1
-
RECOMMENDED NAME
GeneOntology No.
ferroxidase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
4 Fe(II) + 4 H+ + O2 = 4 Fe(III) + 2 H2O
show the reaction diagram
steady state kinetic analysis, ferroxidase and cuprous oxidase activities are due to the same electron transfer site on the enzyme
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
non-pathway related
-
-
Porphyrin and chlorophyll metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
Fe(II):oxygen oxidoreductase
The enzyme in blood plasma (ceruloplasmin) belongs to the family of multicopper oxidases. In humans it accounts for 95% of plasma copper. It oxidizes Fe(II) to Fe(III), which allows the subsequent incorporation of the latter into proteins such as apotransferrin and lactoferrin. An enzyme from iron oxidizing bacterium strain TI-1 contains heme a.
CAS REGISTRY NUMBER
COMMENTARY hide
9031-37-2
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
additional information
ferritin is a ubiquitous iron storage protein that possesses ferroxidase activity
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2-chloro-p-phenylenediamine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
2-methoxy-p-phenylenediamine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
2-methyl-p-phenylenediamine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
2-nitro-p-phenylenediamine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
2-sulfonic acid-p-phenylenediamine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
3,4-dihydroxyphenethylamine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
4 Cu+ + 4 H+ + O2
4 Cu2+ + 2 H2O
show the reaction diagram
-
-
-
-
?
4 Fe(II) + 4 H+ + O2
4 Fe(III) + 2 H2O
show the reaction diagram
-
-
-
-
?
4-methylcatechol + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
4-phenylenediamine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
5-hydroxyindol-3-ylacetic acid + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
5-hydroxytryptamine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
5-hydroxytryptophan + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
5-hydroxytryptophol + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
alimemazine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
apotransferrin + Fe2+
holotransferrin + ?
show the reaction diagram
-
-
-
-
?
apotransferrin + Fe2+ + O2
diferric transferrin + H2O
show the reaction diagram
-
-
-
-
?
ascorbate + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
catechol + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
chlorpromazine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
diethazine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
dihydroxyphenylethylamine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
durenediamine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
Fe2+ + H+ + O2
Fe3+ + H2O
show the reaction diagram
ferrous ammonium sulfate + O2
?
show the reaction diagram
-
-
-
-
?
ferrous ammonium sulfate + O2
? + H2O
show the reaction diagram
-
-
-
-
?
fluphenazine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
L-epinephrine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
L-norepinephrine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
m-phenylenediamine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
N,N'-dimethyl-p-phenylenediamine + Cu2+ + O2
N,N'-dimethyl-p-phenylenediamine radical + Cu+
show the reaction diagram
-
-
-
-
?
N,N'-dimethyl-p-phenylenediamine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
N,N'-dimethyl-p-phenylenediamine + Fe3+ + O2
?
show the reaction diagram
-
-
-
-
?
N,N,N',N'-tetramethyl-p-phenylenediamine + Fe2+
?
show the reaction diagram
-
-
-
-
?
N,N-diethyl-p-phenylenediamine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
N,N-dimethyl-m-phenylenediamine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
N,N-dimethyl-p-phenylenediamine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
N-(p-methoxyphenyl)-p-phenylenediamine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
N-acetyl-p-phenylenediamine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
N-ethyl-N-(2-hydroxyethyl)-p-phenylenediamine Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
N-ethyl-N-2(S-methylsulfonamido)-ethyl-p-phenylenediamine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
N-phenyl-p-phenylenediamine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
o-aminophenol + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
o-dianisidine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
o-phenylenediamine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
p-aminophenol + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
p-anisidine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
p-phenylenediamine + Cu2+ + O2
p-phenylenediamine radical + Cu+
show the reaction diagram
p-phenylenediamine + Fe2+ + O2
?
show the reaction diagram
periciazine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
perphenazine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
prochlorperazine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
promazine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
prometazine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
pyrogallol + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
quinone + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
thioridazine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
trifluoperazine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
triflupromazine + Fe2+ + O2
?
show the reaction diagram
-
-
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
4 Fe(II) + 4 H+ + O2
4 Fe(III) + 2 H2O
show the reaction diagram
-
-
-
-
?
Fe2+ + H+ + O2
Fe3+ + H2O
show the reaction diagram
additional information
?
-
-
treatment of mouse BV-2 cells and primary microglial cells with ceruloplasmin induces nitric oxide release and inducible NO synthase mRNA expression. Presence of ceruloplasmin increases levels of mRNAs encoding tumor necrosis factor-alpha, interleukin-1beta, cyclooxygenase-2, and NADPH oxidase. Treatment of BV-2 cells and primary microglia with ceruloplasmin induces phosphorylation of p38 MAP kinase. Ceruloplasmin induces nuclear factor kappaB activation, showing a more sustained pattern than seen with bacterial lipopolysaccharide. Ceruloplasmin-stimulated NO induction is significantly attenuated by p38 inhibitor, SB203580, and the nucleare factor kappaB inhibitor SN50. Ceruloplasmin induces secretion of tumor necrosis factor-alpha and prostaglandin E2 in primary microglial cultures
-
-
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
copper
-
3.1 atoms per protein molecule
Cr3+
-
0.1 M, activity 102%
Iron
-
ceruloplasmin is regulated by cellular iron status
Ni2+
-
activating
Zn2+
-
activating
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
6-aminohexanoic acid
-
-
Ba2+
-
weak inhibitor
Ca2+
-
weak inhibitor
Cr3+
-
strong inhibitor
K+
-
weak inhibitor
Li+
-
weak inhibitor
N3-
-
anion behaves as an inhibitor of the oxidase activity versus Fe2+
Na+
-
weak inhibitor
Sn2+
-
weak inhibitor
Sodium azide
Zn2+
-
2 M Zn2+ occupies the ferroxidase center as redox-invariant analogue of Fe2+
ZrO2+
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
lactoferrin
-
apolactoferrin increases the oxidation rate of Fe(II) by ceruloplasmin 1.25fold at pH 5.5. Lactoferrin saturated with Fe3+ or Cu2+ increases the oxidation rate of Fe2+ 1.6fold when in 1:1 ratio with enzyme
-
pyrrolidine dithiocarbamate
-
regulation of ceruloplasmin expression by a copper-dependent transcriptional mechanism
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.241
2-chloro-p-phenylenediamine
-
-
0.161
2-methoxy-p-phenylenediamine
-
-
0.213
2-methyl-p-phenylenediamine
-
-
0.00126
2-nitro-p-phenylenediamine
-
-
0.00262
2-sulfonic acid-p-phenylenediamine
-
-
0.00285
3,4-Dihydroxyphenethylamine
-
-
0.0603
4-methylcatechol
-
-
0.0015
4-phenylenediamine
-
pH 5, 20C
0.00834
5-hydroxyindol-3-ylacetic acid
-
-
0.908
5-hydroxytryptamine
-
-
0.0163
5-hydroxytryptophan
-
-
0.0051
5-hydroxytryptophol
-
-
0.0014
alimemazine
-
-
0.0052
ascorbate
-
-
0.282
catechol
-
-
0.0035
chlorpromazine
-
-
0.0368
Cu+
-
pH 5.0
0.0023
diethazine
-
-
0.171
durenediamine
-
-
0.0006 - 0.05
Fe2+
0.0021
Ferrous ammonium sulfate
-
pH 5.0, 22C
0.005
fluphenazine
-
-
0.00255
L-epinephrine
-
-
0.00281
L-norepinephrine
-
-
0.036
m-phenylenediamine
-
-
0.11 - 0.164
N,N'-dimethyl-p-phenylenediamine
0.197
N,N,N',N'-tetramethyl-p-phenylenediamine
-
-
0.556
N,N-diethyl-p-phenylenediamine
-
-
0.00305
N,N-dimethyl-m-phenylenediamine
-
-
0.203
N,N-Dimethyl-p-phenylenediamine
-
-
0.021
N-(p-methoxyphenyl)p-phenylenediamine
-
-
0.0123
N-acetyl-p-phenylenediamine
-
-
0.11
N-ethyl-N-(2-hydroxyethyl)p-phenylenediamine
-
-
0.087
N-ethyl-N-2(S-methylsulfonamido)-ethyl-p-phenylenediamine
-
-
0.048
N-phenyl-p-phenylenediamine
-
-
0.00288
o-aminophenol
-
-
0.12 - 0.18
o-Dianisidine
0.00295
o-phenylenediamine
-
-
0.0074 - 0.0632
O2
0.00154
p-Aminophenol
-
-
0.00614
p-anisidine
-
-
0.292 - 2.5
p-phenylenediamine
0.002
periciazine
-
-
0.0013
perphenazine
-
-
0.9
prochlorperazine
-
-
0.0013
promazine
-
-
0.0023
Promethazine
-
-
0.0579
pyrogallol
-
-
0.0657
quinone
-
-
0.0014
thioridazine
-
-
0.0028
Trifluoperazine
-
-
0.01
triflupromazine
-
-
additional information
additional information
stopped-flow kinetics and initial velocities of iron oxidation of wild-type and mutant enzymes, overview
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.48
4-phenylenediamine
-
pH 5, 20C
0.375
Cu+
-
pH 5.0
0.3 - 0.505
Fe2+
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.28
-
-
0.58
-
in an Erel assay measuring loss of Fe2+
0.88
-
o-dianisidine as substrate
1.25
-
in a transferrin assay measuring transformation of apotransferrin to holotransferrin
1.73
-
p-phenylenediamine as substrate
3.13
-
-
30
-
ferroxidase II
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 5.2
-
schizophrenics
5.5 - 5.6
-
normal persons
5.7
-
phosphate buffer
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 7.5
-
-
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
assay at
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
in fluid from patients with Parkinson's disease, Alzheimer's disease, Huntington's disease, a significantly decreased ferroxidase activity is found agreeing with findings of iron deposition in these entities, while free copper is found to be increased in cerebrospinal fluid and appears to be a good biomarker of Parkinson's disease. The sum of nitrites and nitrates as end products of nitric oxide are increased in the degenerative diseases Parkinson's disease, Alzheimer's disease, Huntington's disease and lateral amyotrophic sclerosis and fluorescent lipoperoxidation products in three of them, excepting lateral amyotrophic sclerosis
Manually annotated by BRENDA team
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nonadherent cells, T and B lymphocytes
Manually annotated by BRENDA team
-
-
Manually annotated by BRENDA team
-
nephrotic urine
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
UNIPROT
ORGANISM
Homo sapiens;
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
15900
-
2 * 15900 + 2 * 58900, SDS-PAGE
17000
-
1 * 17000 + 1 * 59000
18650
-
1 * 18650 + 1 * 50000 + 1 * 70000, limited proteolysis
19000
-
1 * 19000 + 1 * 25000 + 1 * 26000 + 1 * 67000, SDS-PAGE; 1 * 19000 + 1 * 50000 + 1 * 67000, limited proteolysis
24000
-
1 * 24000 + 1 * 93000, cleaved by protease
25000
-
1 * 19000 + 1 * 25000 + 1 * 26000 + 1 * 67000, SDS-PAGE
26000
-
1 * 19000 + 1 * 25000 + 1 * 26000 + 1 * 67000, SDS-PAGE
35000
-
2 * 16000 + 2 * 35000, SDS-PAGE
58900
-
2 * 15900 + 2 * 58900, SDS-PAGE
67000
-
1 * 19000 + 1 * 25000 + 1 * 26000 + 1 * 67000, SDS-PAGE; 1 * 19000 + 1 * 50000 + 1 * 67000, limited proteolysis
70000
-
1 * 18650 + 1 * 50000 + 1 * 70000, limited proteolysis
93000
-
1 * 24000 + 1 * 93000, cleaved by protease
100000 - 200000
-
gel filtration
115000
-
human ceruloplasmin antibody reaction
120000
121000
124000
129000
-
determination of peptide chain length
129600
-
x * 129600, MALDI-TOF of recombinant glycoprotein, x * 143000, SDS-PAGE of recombinant protein, x * 121000, SDS-PAGE of PNGase F-treated protein
130000
131000
-
sedimentation equilibrium
132000
134000
135000
137000
-
gel filtration
143000
-
x * 129600, MALDI-TOF of recombinant glycoprotein, x * 143000, SDS-PAGE of recombinant protein, x * 121000, SDS-PAGE of PNGase F-treated protein
155000
-
sedimentation equilibrium
160000
200000
-
human ceruloplasmin antibody reaction
800000 - 2000000
-
ferroxidase II, gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
24-mer
-
advanced protein assay
dimer
-
1 * 17000 + 1 * 59000; 1 * 24000 + 1 * 93000, cleaved by protease
tetramer
trimer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified recombinant H ferritin, 10 mg/ml protein in 10 mM Tris,HCl, pH 7.5, and 0.15 M NaCl, is mixed at equal volumes with crystallization solution containing 0.1 M BICINE-Na, pH 9.0, and 1.9-2.0 M MgCl2, 20C, 1 week, X-ray diffraction structure determination and analysis at 1.52-1.97 A resolution
small angel X-ray scattering analysis of the complex with lactoferrin. Ceruloplasmin forms a 1:1 complex with lactoferrin. Complex formation occurs without major conformational rearrangements of either protein
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.8 - 6.4
-
above pH 5.8 activity progressively decreases
437996
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
65
-
above irreversible denaturation process of the protein active site
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 0.5 M phosphate buffer, pH 6.9, concentration of approximately 2.5% without noticeable loss of blue colour during 1 month
-
-20C, A280:A610 ratio increases for several weeks on storage
-
-20C, frozen in liquid nitrogen 0.2% enzyme solution in 0.015 M phosphate buffer, pH 6.9, containing 0.1 M NaCl loses its blue colour completely after 2 weeks storage
-
-90C, when frozen in dry ice and thawed, about 5% of the absorbance at 610 nm is lost, no change in the absorbance at 280 nm
-
4C, 0.1 M Tris buffer, pH 8.0, decomposes into fragments when stored for 36-48 h
-
4C, 0.2% enzyme solution in 0.015 M phosphate buffer, pH 6.9, containing 0.1 M NaCl: the A610/A280 ratio is reduced by approximately 10% in 1 month
-
4C, ferroxidase II, purified enzyme is stable for at least 2 weeks
-
4C, purified enzyme sensitive to storage, no oxidase activity after 4 months
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
after expression in baby hamster kidney cells
-
blood plasma ceruloplasmin was purified by DEAE-Sepharose chromatography
-
non-ceruloplasmin ferroxidase II
-
partially
-
recombinant ceruloplasmin, expressed in Pichia pastoris GS115 his4
-
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by heat treatment at 60C for 10 min, followed by ammonium sulfate fractionation, anion exchange chromatography, and gel filtration
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cDNA clones encoding human CP identified, CP gene mapped to human chromosome 3q21-25 by human-mouse somatic-cell-hybrid analysis
-
expressed in Escherichia coli
-
expressed in expressed in baby hamster kidney cells
-
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
fully active recombinant human ceruloplasmin produced in the yeast Pichia pastoris
-
gene sequencing and site-directed mutagenesis
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D616A/H621A/E960A/H965A
-
iron binding sites are mutated but one is unaffected. Km and kcat for substrate 4-phenylenediamine decreased compared to wild-type. kcat (Fe2+) up to 10fold decreased compared to wild-type. Mutant does not retain a high-affinity iron oxidation component
E140A
site-directed mutagenesis, the initial velocity of iron oxidization is reduced in the mutant
E140Q
site-directed mutagenesis, the initial velocity of iron oxidization is highly reduced in the mutant. The side chain of the mutated Gln140 is fixed by a hydrogen bond, whereas that of native Glu140 is flexible
E264A/H269A/D616A/H621A
-
iron binding sites are mutated but one is unaffected. Km and kcat for substrate 4-phenylenediamine decreased compared to wild-type. Km for high-affinity oxidation of Fe2+ decreased compared to wild-type. kcat (Fe2+) up to 10fold decreased compared to wild-type. Only mutant that retains a high-affinity iron oxidation component
E264A/H269A/D616A/H621A/E960A/H965A
-
all three iron binding sites are mutated. Km and kcat for substrate 4-phenylenediamine decreased compared to wild-type. kcat (Fe2+) up to 75 fold decreased compared to wild-type. Mutant does not retain a high-affinity iron oxidation component
E264A/H269A/E960A/H965A
-
iron binding sites are mutated but one is unaffected. Km and kcat for substrate 4-phenylenediamine decreased compared to wild-type. kcat (Fe2+) up to 10fold decreased compared to wild-type. Mutant does not retain a high-affinity iron oxidation component
K86Q
-
equivalent to wild-type
K86Q/E107D
-
reduced reduction activity
K86Q/E27D
-
in X-ray absorption same properties as wild-type, but reduced reduction activity
K86Q/E27D/E107D
-
no reduction activity
W93F/Y34W
-
no alteration in the rate of Fe2+ oxidation
W93F/Y34W/Y29Q
-
no alteration in the rate of Fe2+ oxidation
Y34W/W93F/D131I/E134F
-
no transport of Fe2+ to the ferroxidase center
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine