BRENDA - Enzyme Database show

Molecular characterization of a novel type of prostamide/prostaglandin F synthase, belonging to the thioredoxin-like superfamily

Moriuchi, H.; Koda, N.; Okuda-Ashitaka, E.; Daiyasu, H.; Ogasawara, K.; Toh, H.; Ito, S.; Woodward, D.F.; Watanabe, K.; J. Biol. Chem. 283, 792-801 (2008)

Data extracted from this reference:

Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
1.1.1.188
ammonium sulfate
activity of the purified recombinant enzyme is stimulated about 4fold by 1.5 M
Sus scrofa
1.11.1.20
(NH4)2SO4
stimulates
Sus scrofa
1.11.1.20
ammonium sulfate
the activity of the purified recombinant enzyme is stimulated about 4fold by 1.5 M ammonium sulfate
Mus musculus
Cloned(Commentary)
EC Number
Commentary
Organism
1.1.1.188
expressed in Escherichia coli BL21 cells
Sus scrofa
1.1.1.188
wild-type and mutants, plasmid pCold II-PM/PGFS expressed in Escherichia coli BL21 cells
Mus musculus
1.11.1.20
construction of an expression plasmid using polyhistidine-tagged peptide-fused vector and mouse cDNA encoding a 201-amino acid polypeptide with molecular weight of 21669 Da and expression the protein in Escherichia coli; expressed in Escherichia coli BL21(DE3) cells
Mus musculus
Engineering
EC Number
Amino acid exchange
Commentary
Organism
1.1.1.188
C44S
retains 63% of the enzyme activity of the wild-type
Mus musculus
1.1.1.188
C44S/C47S
shows values below 1% of the enzyme activity of the wild-type
Mus musculus
1.1.1.188
C47S
shows values below 1% of the enzyme activity of the wild-type
Mus musculus
1.11.1.20
C44S
below 1% of wild-type activity; the mutant is almost inactive with less than 1% of wild type activity
Mus musculus
1.11.1.20
C44S/C47S
below 1% of wild-type activity; the mutant is almost inactive with less than 1% of wild type activity
Mus musculus
1.11.1.20
C47S
mutant shows reduced activity compared to the wild type enzyme; retains 63% of wild-type activity
Mus musculus
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
1.11.1.20
cumene hydroperoxide
competitive inhibition of prostamide F synthase activity
Mus musculus
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.188
0.0069
-
prostaglandin H2
pH 7.0, 24°C
Mus musculus
1.1.1.188
0.007
-
prostaglandin H2
-
Sus scrofa
1.11.1.20
0.001
-
thioredoxin
pH 7.0, 24°C
Mus musculus
1.11.1.20
0.0069
-
(5Z,9alpha,11alpha,13E,15S)-9,11-epidioxy-15-hydroxy-prosta-5,13-dienoate
pH 7.0, 24°C, i.e. prostaglandin H2
Mus musculus
1.11.1.20
0.0069
-
H2O2
in 0.1 M KPB (pH 7.0), at 24°C
Mus musculus
1.11.1.20
0.0069
-
prostaglandin H2
in 0.1 M KPB (pH 7.0), at 24°C
Mus musculus
1.11.1.20
0.007
-
(5Z,9alpha,11alpha,13E,15S)-9,11-epidioxy-15-hydroxy-prosta-5,13-dienoate
i.e. prostaglandin H2, pH 7.0, 24°C
Sus scrofa
1.11.1.20
0.0076
-
(5Z)-N-(2-hydroxyethyl)-7-[(1R,4S,5R,6R)-6-[(1E,3S)-3-hydroxyoct-1-en-1-yl]-2,3-dioxabicyclo[2.2.1]hept-5-yl]hept-5-enamide
pH 7.0, 24°C, i.e. prostamide H2
Mus musculus
1.11.1.20
0.0076
-
prostamide H2
in 0.1 M KPB (pH 7.0), at 24°C
Mus musculus
1.11.1.20
0.014
-
(5Z)-N-(2-hydroxyethyl)-7-[(1R,4S,5R,6R)-6-[(1E,3S)-3-hydroxyoct-1-en-1-yl]-2,3-dioxabicyclo[2.2.1]hept-5-yl]hept-5-enamide
i.e. prostamide H2, pH 7.0, 24°C
Sus scrofa
1.11.1.20
0.026
-
cumene hydroperoxide
in 0.1 M KPB (pH 7.0), at 24°C; pH 7.0, 24°C
Mus musculus
1.11.1.20
0.4
-
t-butyl hydroperoxide
in 0.1 M KPB (pH 7.0), at 24°C
Mus musculus
1.11.1.20
0.4
-
tert-butyl hydroperoxide
pH 7.0, 24°C
Mus musculus
1.11.1.20
1.6
-
H2O2
pH 7.0, 24°C
Mus musculus
Localization
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
1.1.1.188
cytoplasm
-
Mus musculus
5737
-
1.1.1.188
endoplasmic reticulum
-
Mus musculus
5783
-
1.11.1.20
cytosol
-
Mus musculus
5829
-
1.11.1.20
endoplasmic reticulum
-
Mus musculus
5783
-
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
1.11.1.20
(NH4)2SO4
1.5 M, stimulates about 4fold
Mus musculus
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.1.1.188
20000
-
immunoprecipitation, Western blot analysis
Sus scrofa
1.1.1.188
23000
-
by zinc staining
Mus musculus
1.1.1.188
23230
-
expressed protein
Mus musculus
1.11.1.20
20000
-
SDS-PAGE
Mus musculus
1.11.1.20
20000
-
x * 20000, SDS-PAGE
Sus scrofa
1.11.1.20
21670
-
calculated from amino acid sequence
Mus musculus
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.1.1.188
Mus musculus
-
-
-
1.1.1.188
Sus scrofa
A9CQL8
-
-
1.11.1.20
Mus musculus
Q9DB60
-
-
1.11.1.20
Sus scrofa
A9CQL8
-
-
Purification (Commentary)
EC Number
Commentary
Organism
1.1.1.188
on Ni2+ column, 102- and 152fold purification of the prostamide F synthase and PGF synthase activities, respectively, achieved from the swine brain (90 g) with a yield of 1.4 and 2%, respectively. Mutants expressed to homogeneity
Sus scrofa
1.1.1.188
to apparent homogeneity on Ni2+ column
Mus musculus
1.11.1.20
-
Sus scrofa
1.11.1.20
; Ni2+-agarose column chromatography and Q Sepharose column chromatography
Mus musculus
Source Tissue
EC Number
Source Tissue
Commentary
Organism
Textmining
1.1.1.188
adrenal gland
-
Mus musculus
-
1.1.1.188
brain
high activity
Mus musculus
-
1.1.1.188
brain
highest activity
Sus scrofa
-
1.1.1.188
cecum
low activity
Mus musculus
-
1.1.1.188
eye
low activity
Mus musculus
-
1.1.1.188
eye
-
Sus scrofa
-
1.1.1.188
heart
high activity
Mus musculus
-
1.1.1.188
heart
-
Sus scrofa
-
1.1.1.188
kidney
low activity
Mus musculus
-
1.1.1.188
large intestine
low activity
Mus musculus
-
1.1.1.188
liver
low activity
Mus musculus
-
1.1.1.188
lung
low activity
Mus musculus
-
1.1.1.188
lung
-
Sus scrofa
-
1.1.1.188
additional information
low activity in deferent duct and epididymidis
Mus musculus
-
1.1.1.188
muscle
low activity
Mus musculus
-
1.1.1.188
ovary
high activity
Mus musculus
-
1.1.1.188
small intestine
low activity
Mus musculus
-
1.1.1.188
spinal cord
-
Mus musculus
-
1.1.1.188
spleen
low activity
Mus musculus
-
1.1.1.188
stomach
low activity
Mus musculus
-
1.1.1.188
testis
low activity
Mus musculus
-
1.1.1.188
thymus
high activity
Mus musculus
-
1.1.1.188
uterus
high activity
Mus musculus
-
1.1.1.188
vesicular gland
low activity
Mus musculus
-
1.11.1.20
adrenal gland
; activity is the highest in spinal cord and brain followed by thymus, adrenal gland, heart, and genital organs
Mus musculus
-
1.11.1.20
brain
activity is the highest in spinal cord and brain followed by thymus, adrenal gland, heart, and genital organs; highest activity in the spinal cord and brain
Mus musculus
-
1.11.1.20
brain
-
Sus scrofa
-
1.11.1.20
cecum
lowest activity in the cecum
Mus musculus
-
1.11.1.20
epididymis
-
Mus musculus
-
1.11.1.20
heart
; activity is the highest in spinal cord and brain followed by thymus, adrenal gland, heart, and genital organs
Mus musculus
-
1.11.1.20
kidney
-
Mus musculus
-
1.11.1.20
large intestine
-
Mus musculus
-
1.11.1.20
liver
-
Mus musculus
-
1.11.1.20
lung
-
Mus musculus
-
1.11.1.20
ovary
; activity is the highest in spinal cord and brain followed by thymus, adrenal gland, heart, and genital organs
Mus musculus
-
1.11.1.20
spinal cord
highest activity in the spinal cord and brain; localized in the superficial layer of the dorsal horn, in motor neurons of the ventral horn, and in glia of the white matter of the spinal cord. Activity is the highest in spinal cord and brain followed by thymus, adrenal gland, heart, and genital organs
Mus musculus
-
1.11.1.20
stomach
-
Mus musculus
-
1.11.1.20
testis
; activity is the highest in spinal cord and brain followed by thymus, adrenal gland, heart, and genital organs
Mus musculus
-
1.11.1.20
thymus
; activity is the highest in spinal cord and brain followed by thymus, adrenal gland, heart, and genital organs
Mus musculus
-
1.11.1.20
uterus
; activity is the highest in spinal cord and brain followed by thymus, adrenal gland, heart, and genital organs
Mus musculus
-
1.11.1.20
vas deferens
-
Mus musculus
-
1.11.1.20
vesicular gland
; activity is the highest in spinal cord and brain followed by thymus, adrenal gland, heart, and genital organs
Mus musculus
-
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.1.1.188
0.062
-
PGF synthase activity of the purified enzyme
Sus scrofa
1.1.1.188
0.69
-
PGF synthase activity of the purified enzyme
Mus musculus
1.11.1.20
0.053
-
prostamine F synthase activity, pH 7.0, 24°C
Sus scrofa
1.11.1.20
0.062
-
prostaglandin F synthase activity, pH 7.0, 24°C
Sus scrofa
1.11.1.20
0.25
-
prostamine F synthase activity, pH 7.0, 24°C; substrate: tert-butyl hydroperoxide, pH 7.0, 24°C
Mus musculus
1.11.1.20
0.28
-
substrate: cumene hydroperoxide, pH 7.0, 24°C; substrate: H2O2, pH 7.0, 24°C
Mus musculus
1.11.1.20
0.69
-
prostaglandin F synthase activity, pH 7.0, 24°C
Mus musculus
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.1.1.188
additional information
no activity with prostaglandin D2 and prostaglandin E2
687797
Mus musculus
?
-
-
-
-
1.1.1.188
prostaglandin H2 + NADPH + H+
-
687797
Mus musculus
prostaglandin F2alpha + NADP+
-
-
-
?
1.1.1.188
prostaglandin H2 + NADPH + H+
-
687797
Sus scrofa
prostaglandin F2alpha + NADP+
-
-
-
?
1.11.1.20
additional information
prostaglandin D2 and prostaglandin E2 are not reduced
687797
Mus musculus
?
-
-
-
-
1.11.1.20
additional information
neither prostaglandin E2 nor prostamide D2 serve as a substrate even at the concentration of 1.5 mM
687797
Mus musculus
?
-
-
-
-
1.11.1.20
additional information
the recombinant enzyme requires thioredoxin as a reducing equivalent donor more efficiently than NADPH, NADH, or GSH
687797
Mus musculus
?
-
-
-
-
1.11.1.20
thioredoxin + (5Z)-N-(2-hydroxyethyl)-7-[(1R,4S,5R,6R)-6-[(1E,3S)-3-hydroxyoct-1-en-1-yl]-2,3-dioxabicyclo[2.2.1]hept-5-yl]hept-5-enamide
(5Z)-N-(2-hydroxyethyl)-7-[(1R,4S,5R,6R)-6-[(1E,3S)-3-hydroxyoct-1-en-1-yl]-2,3-dioxabicyclo[2.2.1]hept-5-yl]hept-5-enamide i.e. prostamide H2
687797
Sus scrofa
thioredoxin disulfide + (5Z,9alpha,11alpha,13E,15S)-9,11,15-trihydroxy-N-(2-hydroxyethyl)prosta-5,13-dien-1-amide
(5Z,9alpha,11alpha,13E,15S)-9,11,15-trihydroxy-N-(2-hydroxyethyl)prosta-5,13-dien-1-amide i.e. prostamide F2alpha
-
-
?
1.11.1.20
thioredoxin + (5Z)-N-(2-hydroxyethyl)-7-[(1R,4S,5R,6R)-6-[(1E,3S)-3-hydroxyoct-1-en-1-yl]-2,3-dioxabicyclo[2.2.1]hept-5-yl]hept-5-enamide
(5Z)-N-(2-hydroxyethyl)-7-[(1R,4S,5R,6R)-6-[(1E,3S)-3-hydroxyoct-1-en-1-yl]-2,3-dioxabicyclo[2.2.1]hept-5-yl]hept-5-enamide i.e. prostamide H2. Prostaglandin H2 and prostamide H2 are the best substrates
687797
Mus musculus
thioredoxin disulfide + (5Z,9alpha,11alpha,13E,15S)-9,11,15-trihydroxy-N-(2-hydroxyethyl)prosta-5,13-dien-1-amide
(5Z,9alpha,11alpha,13E,15S)-9,11,15-trihydroxy-N-(2-hydroxyethyl)prosta-5,13-dien-1-amide i.e. prostamide F2alpha
-
-
?
1.11.1.20
thioredoxin + (5Z,9alpha,11alpha,13E,15S)-9,11-epidioxy-15-hydroxy-prosta-5,13-dienoate
(5Z,9alpha,11alpha,13E,15S)-9,11-epidioxy-15-hydroxy-prosta-5,13-dienoate i.e. grostaglandin H2
687797
Sus scrofa
thioredoxin disulfide + (5Z,9alpha,11alpha,13E,15S)-9,11,15-trihydroxyprosta-5,13-dienoate
(5Z,9alpha,11alpha,13E,15S)-9,11,15-trihydroxyprosta-5,13-dienoate i.e. prostaglandin F2alpha
-
-
?
1.11.1.20
thioredoxin + (5Z,9alpha,11alpha,13E,15S)-9,11-epidioxy-15-hydroxy-prosta-5,13-dienoate
(5Z,9alpha,11alpha,13E,15S)-9,11-epidioxy-15-hydroxy-prosta-5,13-dienoate i.e. prostaglandin H2. Prostaglandin H2 and prostamide H2 are the best substrates. Cys44 is essential for the catalytic activity of prostamide/PGF synthase
687797
Mus musculus
thioredoxin disulfide + (5Z,9alpha,11alpha,13E,15S)-9,11,15-trihydroxyprosta-5,13-dienoate
(5Z,9alpha,11alpha,13E,15S)-9,11,15-trihydroxyprosta-5,13-dienoate i.e. prostaglandin F2alpha
-
-
?
1.11.1.20
thioredoxin + cumene hydroperoxide
-
687797
Mus musculus
thioredoxin disulfide + H2O + 2-phenylpropan-2-ol
-
-
-
?
1.11.1.20
thioredoxin + cumene hydroperoxide
-
687797
Mus musculus
thioredoxin disulfide + ?
-
-
-
?
1.11.1.20
thioredoxin + H2O2
-
687797
Mus musculus
thioredoxin disulfide + 2 H2O
-
-
-
?
1.11.1.20
thioredoxin + H2O2
-
687797
Mus musculus
thioredoxin disulfide + ?
-
-
-
?
1.11.1.20
thioredoxin + prostaglandin H2
prostaglandin H2 is one of the best substrates
687797
Mus musculus
thioredoxin disulfide + prostaglandin F2alpha
-
-
-
?
1.11.1.20
thioredoxin + prostamide H2
prostamide H2 is one of the best substrates
687797
Mus musculus
thioredoxin disulfide + prostamide F2alpha
-
-
-
?
1.11.1.20
thioredoxin + t-butyl hydroperoxide
-
687797
Mus musculus
thioredoxin disulfide + ?
-
-
-
?
1.11.1.20
thioredoxin + tert-butyl hydroperoxide
-
687797
Mus musculus
thioredoxin disulfide + H2O + tert-butanol
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
1.1.1.188
dimer
2 * 23000, wild-type and mutants, SDS-PAGE
Mus musculus
1.1.1.188
dimer
-
Sus scrofa
1.11.1.20
?
x * 21670, stimulation by ammonium sulfate suggests that the enzyme may be a dimer or oligomer instead of a monomer, calculated from sequence
Mus musculus
1.11.1.20
?
x * 20000, SDS-PAGE
Sus scrofa
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.11.1.20
24
-
assay at
Mus musculus
1.11.1.20
24
-
assay at
Sus scrofa
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.1.1.188
0.2467
-
prostaglandin H2
pH 7.0, 24°C
Mus musculus
1.1.1.188
0.2467
-
prostaglandin H2
-
Sus scrofa
1.11.1.20
0.001
-
thioredoxin
in 0.1 M KPB (pH 7.0), at 24°C
Mus musculus
1.11.1.20
0.09
-
(5Z)-N-(2-hydroxyethyl)-7-[(1R,4S,5R,6R)-6-[(1E,3S)-3-hydroxyoct-1-en-1-yl]-2,3-dioxabicyclo[2.2.1]hept-5-yl]hept-5-enamide
pH 7.0, 24°C, i.e. prostamide H2
Mus musculus
1.11.1.20
0.09
-
prostamide H2
in 0.1 M KPB (pH 7.0), at 24°C
Mus musculus
1.11.1.20
0.09
-
t-butyl hydroperoxide
in 0.1 M KPB (pH 7.0), at 24°C
Mus musculus
1.11.1.20
0.09
-
tert-butyl hydroperoxide
pH 7.0, 24°C
Mus musculus
1.11.1.20
0.1
-
cumene hydroperoxide
in 0.1 M KPB (pH 7.0), at 24°C; pH 7.0, 24°C
Mus musculus
1.11.1.20
0.1
-
H2O2
in 0.1 M KPB (pH 7.0), at 24°C; pH 7.0, 24°C
Mus musculus
1.11.1.20
0.25
-
(5Z,9alpha,11alpha,13E,15S)-9,11-epidioxy-15-hydroxy-prosta-5,13-dienoate
pH 7.0, 24°C, i.e. prostaglandin H2
Mus musculus
1.11.1.20
0.25
-
prostaglandin H2
in 0.1 M KPB (pH 7.0), at 24°C
Mus musculus
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.11.1.20
7
-
; optimal pH for the prostamide F synthase activity
Mus musculus
1.11.1.20
7
-
assay at
Sus scrofa
pH Range
EC Number
pH Minimum
pH Maximum
Commentary
Organism
1.11.1.20
6.5
8
pH 6.5: about 50% of maximal activity, pH 8.0: about 70% of maximal activity
Mus musculus
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.188
NADPH
-
Sus scrofa
1.1.1.188
NADPH
-
Mus musculus
1.11.1.20
thioredoxin
-
Mus musculus
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
1.1.1.188
ammonium sulfate
activity of the purified recombinant enzyme is stimulated about 4fold by 1.5 M
Sus scrofa
1.11.1.20
(NH4)2SO4
stimulates
Sus scrofa
1.11.1.20
ammonium sulfate
the activity of the purified recombinant enzyme is stimulated about 4fold by 1.5 M ammonium sulfate
Mus musculus
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.188
expressed in Escherichia coli BL21 cells
Sus scrofa
1.1.1.188
wild-type and mutants, plasmid pCold II-PM/PGFS expressed in Escherichia coli BL21 cells
Mus musculus
1.11.1.20
construction of an expression plasmid using polyhistidine-tagged peptide-fused vector and mouse cDNA encoding a 201-amino acid polypeptide with molecular weight of 21669 Da and expression the protein in Escherichia coli; expressed in Escherichia coli BL21(DE3) cells
Mus musculus
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.188
NADPH
-
Sus scrofa
1.1.1.188
NADPH
-
Mus musculus
1.11.1.20
thioredoxin
-
Mus musculus
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
1.1.1.188
C44S
retains 63% of the enzyme activity of the wild-type
Mus musculus
1.1.1.188
C44S/C47S
shows values below 1% of the enzyme activity of the wild-type
Mus musculus
1.1.1.188
C47S
shows values below 1% of the enzyme activity of the wild-type
Mus musculus
1.11.1.20
C44S
below 1% of wild-type activity; the mutant is almost inactive with less than 1% of wild type activity
Mus musculus
1.11.1.20
C44S/C47S
below 1% of wild-type activity; the mutant is almost inactive with less than 1% of wild type activity
Mus musculus
1.11.1.20
C47S
mutant shows reduced activity compared to the wild type enzyme; retains 63% of wild-type activity
Mus musculus
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
1.11.1.20
cumene hydroperoxide
competitive inhibition of prostamide F synthase activity
Mus musculus
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.188
0.0069
-
prostaglandin H2
pH 7.0, 24°C
Mus musculus
1.1.1.188
0.007
-
prostaglandin H2
-
Sus scrofa
1.11.1.20
0.001
-
thioredoxin
pH 7.0, 24°C
Mus musculus
1.11.1.20
0.0069
-
(5Z,9alpha,11alpha,13E,15S)-9,11-epidioxy-15-hydroxy-prosta-5,13-dienoate
pH 7.0, 24°C, i.e. prostaglandin H2
Mus musculus
1.11.1.20
0.0069
-
H2O2
in 0.1 M KPB (pH 7.0), at 24°C
Mus musculus
1.11.1.20
0.0069
-
prostaglandin H2
in 0.1 M KPB (pH 7.0), at 24°C
Mus musculus
1.11.1.20
0.007
-
(5Z,9alpha,11alpha,13E,15S)-9,11-epidioxy-15-hydroxy-prosta-5,13-dienoate
i.e. prostaglandin H2, pH 7.0, 24°C
Sus scrofa
1.11.1.20
0.0076
-
(5Z)-N-(2-hydroxyethyl)-7-[(1R,4S,5R,6R)-6-[(1E,3S)-3-hydroxyoct-1-en-1-yl]-2,3-dioxabicyclo[2.2.1]hept-5-yl]hept-5-enamide
pH 7.0, 24°C, i.e. prostamide H2
Mus musculus
1.11.1.20
0.0076
-
prostamide H2
in 0.1 M KPB (pH 7.0), at 24°C
Mus musculus
1.11.1.20
0.014
-
(5Z)-N-(2-hydroxyethyl)-7-[(1R,4S,5R,6R)-6-[(1E,3S)-3-hydroxyoct-1-en-1-yl]-2,3-dioxabicyclo[2.2.1]hept-5-yl]hept-5-enamide
i.e. prostamide H2, pH 7.0, 24°C
Sus scrofa
1.11.1.20
0.026
-
cumene hydroperoxide
in 0.1 M KPB (pH 7.0), at 24°C; pH 7.0, 24°C
Mus musculus
1.11.1.20
0.4
-
t-butyl hydroperoxide
in 0.1 M KPB (pH 7.0), at 24°C
Mus musculus
1.11.1.20
0.4
-
tert-butyl hydroperoxide
pH 7.0, 24°C
Mus musculus
1.11.1.20
1.6
-
H2O2
pH 7.0, 24°C
Mus musculus
Localization (protein specific)
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
1.1.1.188
cytoplasm
-
Mus musculus
5737
-
1.1.1.188
endoplasmic reticulum
-
Mus musculus
5783
-
1.11.1.20
cytosol
-
Mus musculus
5829
-
1.11.1.20
endoplasmic reticulum
-
Mus musculus
5783
-
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
1.11.1.20
(NH4)2SO4
1.5 M, stimulates about 4fold
Mus musculus
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.1.1.188
20000
-
immunoprecipitation, Western blot analysis
Sus scrofa
1.1.1.188
23000
-
by zinc staining
Mus musculus
1.1.1.188
23230
-
expressed protein
Mus musculus
1.11.1.20
20000
-
SDS-PAGE
Mus musculus
1.11.1.20
20000
-
x * 20000, SDS-PAGE
Sus scrofa
1.11.1.20
21670
-
calculated from amino acid sequence
Mus musculus
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.188
on Ni2+ column, 102- and 152fold purification of the prostamide F synthase and PGF synthase activities, respectively, achieved from the swine brain (90 g) with a yield of 1.4 and 2%, respectively. Mutants expressed to homogeneity
Sus scrofa
1.1.1.188
to apparent homogeneity on Ni2+ column
Mus musculus
1.11.1.20
-
Sus scrofa
1.11.1.20
; Ni2+-agarose column chromatography and Q Sepharose column chromatography
Mus musculus
Source Tissue (protein specific)
EC Number
Source Tissue
Commentary
Organism
Textmining
1.1.1.188
adrenal gland
-
Mus musculus
-
1.1.1.188
brain
high activity
Mus musculus
-
1.1.1.188
brain
highest activity
Sus scrofa
-
1.1.1.188
cecum
low activity
Mus musculus
-
1.1.1.188
eye
low activity
Mus musculus
-
1.1.1.188
eye
-
Sus scrofa
-
1.1.1.188
heart
high activity
Mus musculus
-
1.1.1.188
heart
-
Sus scrofa
-
1.1.1.188
kidney
low activity
Mus musculus
-
1.1.1.188
large intestine
low activity
Mus musculus
-
1.1.1.188
liver
low activity
Mus musculus
-
1.1.1.188
lung
low activity
Mus musculus
-
1.1.1.188
lung
-
Sus scrofa
-
1.1.1.188
additional information
low activity in deferent duct and epididymidis
Mus musculus
-
1.1.1.188
muscle
low activity
Mus musculus
-
1.1.1.188
ovary
high activity
Mus musculus
-
1.1.1.188
small intestine
low activity
Mus musculus
-
1.1.1.188
spinal cord
-
Mus musculus
-
1.1.1.188
spleen
low activity
Mus musculus
-
1.1.1.188
stomach
low activity
Mus musculus
-
1.1.1.188
testis
low activity
Mus musculus
-
1.1.1.188
thymus
high activity
Mus musculus
-
1.1.1.188
uterus
high activity
Mus musculus
-
1.1.1.188
vesicular gland
low activity
Mus musculus
-
1.11.1.20
adrenal gland
; activity is the highest in spinal cord and brain followed by thymus, adrenal gland, heart, and genital organs
Mus musculus
-
1.11.1.20
brain
activity is the highest in spinal cord and brain followed by thymus, adrenal gland, heart, and genital organs; highest activity in the spinal cord and brain
Mus musculus
-
1.11.1.20
brain
-
Sus scrofa
-
1.11.1.20
cecum
lowest activity in the cecum
Mus musculus
-
1.11.1.20
epididymis
-
Mus musculus
-
1.11.1.20
heart
; activity is the highest in spinal cord and brain followed by thymus, adrenal gland, heart, and genital organs
Mus musculus
-
1.11.1.20
kidney
-
Mus musculus
-
1.11.1.20
large intestine
-
Mus musculus
-
1.11.1.20
liver
-
Mus musculus
-
1.11.1.20
lung
-
Mus musculus
-
1.11.1.20
ovary
; activity is the highest in spinal cord and brain followed by thymus, adrenal gland, heart, and genital organs
Mus musculus
-
1.11.1.20
spinal cord
highest activity in the spinal cord and brain; localized in the superficial layer of the dorsal horn, in motor neurons of the ventral horn, and in glia of the white matter of the spinal cord. Activity is the highest in spinal cord and brain followed by thymus, adrenal gland, heart, and genital organs
Mus musculus
-
1.11.1.20
stomach
-
Mus musculus
-
1.11.1.20
testis
; activity is the highest in spinal cord and brain followed by thymus, adrenal gland, heart, and genital organs
Mus musculus
-
1.11.1.20
thymus
; activity is the highest in spinal cord and brain followed by thymus, adrenal gland, heart, and genital organs
Mus musculus
-
1.11.1.20
uterus
; activity is the highest in spinal cord and brain followed by thymus, adrenal gland, heart, and genital organs
Mus musculus
-
1.11.1.20
vas deferens
-
Mus musculus
-
1.11.1.20
vesicular gland
; activity is the highest in spinal cord and brain followed by thymus, adrenal gland, heart, and genital organs
Mus musculus
-
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.1.1.188
0.062
-
PGF synthase activity of the purified enzyme
Sus scrofa
1.1.1.188
0.69
-
PGF synthase activity of the purified enzyme
Mus musculus
1.11.1.20
0.053
-
prostamine F synthase activity, pH 7.0, 24°C
Sus scrofa
1.11.1.20
0.062
-
prostaglandin F synthase activity, pH 7.0, 24°C
Sus scrofa
1.11.1.20
0.25
-
prostamine F synthase activity, pH 7.0, 24°C; substrate: tert-butyl hydroperoxide, pH 7.0, 24°C
Mus musculus
1.11.1.20
0.28
-
substrate: cumene hydroperoxide, pH 7.0, 24°C; substrate: H2O2, pH 7.0, 24°C
Mus musculus
1.11.1.20
0.69
-
prostaglandin F synthase activity, pH 7.0, 24°C
Mus musculus
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.1.1.188
additional information
no activity with prostaglandin D2 and prostaglandin E2
687797
Mus musculus
?
-
-
-
-
1.1.1.188
prostaglandin H2 + NADPH + H+
-
687797
Mus musculus
prostaglandin F2alpha + NADP+
-
-
-
?
1.1.1.188
prostaglandin H2 + NADPH + H+
-
687797
Sus scrofa
prostaglandin F2alpha + NADP+
-
-
-
?
1.11.1.20
additional information
prostaglandin D2 and prostaglandin E2 are not reduced
687797
Mus musculus
?
-
-
-
-
1.11.1.20
additional information
neither prostaglandin E2 nor prostamide D2 serve as a substrate even at the concentration of 1.5 mM
687797
Mus musculus
?
-
-
-
-
1.11.1.20
additional information
the recombinant enzyme requires thioredoxin as a reducing equivalent donor more efficiently than NADPH, NADH, or GSH
687797
Mus musculus
?
-
-
-
-
1.11.1.20
thioredoxin + (5Z)-N-(2-hydroxyethyl)-7-[(1R,4S,5R,6R)-6-[(1E,3S)-3-hydroxyoct-1-en-1-yl]-2,3-dioxabicyclo[2.2.1]hept-5-yl]hept-5-enamide
(5Z)-N-(2-hydroxyethyl)-7-[(1R,4S,5R,6R)-6-[(1E,3S)-3-hydroxyoct-1-en-1-yl]-2,3-dioxabicyclo[2.2.1]hept-5-yl]hept-5-enamide i.e. prostamide H2
687797
Sus scrofa
thioredoxin disulfide + (5Z,9alpha,11alpha,13E,15S)-9,11,15-trihydroxy-N-(2-hydroxyethyl)prosta-5,13-dien-1-amide
(5Z,9alpha,11alpha,13E,15S)-9,11,15-trihydroxy-N-(2-hydroxyethyl)prosta-5,13-dien-1-amide i.e. prostamide F2alpha
-
-
?
1.11.1.20
thioredoxin + (5Z)-N-(2-hydroxyethyl)-7-[(1R,4S,5R,6R)-6-[(1E,3S)-3-hydroxyoct-1-en-1-yl]-2,3-dioxabicyclo[2.2.1]hept-5-yl]hept-5-enamide
(5Z)-N-(2-hydroxyethyl)-7-[(1R,4S,5R,6R)-6-[(1E,3S)-3-hydroxyoct-1-en-1-yl]-2,3-dioxabicyclo[2.2.1]hept-5-yl]hept-5-enamide i.e. prostamide H2. Prostaglandin H2 and prostamide H2 are the best substrates
687797
Mus musculus
thioredoxin disulfide + (5Z,9alpha,11alpha,13E,15S)-9,11,15-trihydroxy-N-(2-hydroxyethyl)prosta-5,13-dien-1-amide
(5Z,9alpha,11alpha,13E,15S)-9,11,15-trihydroxy-N-(2-hydroxyethyl)prosta-5,13-dien-1-amide i.e. prostamide F2alpha
-
-
?
1.11.1.20
thioredoxin + (5Z,9alpha,11alpha,13E,15S)-9,11-epidioxy-15-hydroxy-prosta-5,13-dienoate
(5Z,9alpha,11alpha,13E,15S)-9,11-epidioxy-15-hydroxy-prosta-5,13-dienoate i.e. grostaglandin H2
687797
Sus scrofa
thioredoxin disulfide + (5Z,9alpha,11alpha,13E,15S)-9,11,15-trihydroxyprosta-5,13-dienoate
(5Z,9alpha,11alpha,13E,15S)-9,11,15-trihydroxyprosta-5,13-dienoate i.e. prostaglandin F2alpha
-
-
?
1.11.1.20
thioredoxin + (5Z,9alpha,11alpha,13E,15S)-9,11-epidioxy-15-hydroxy-prosta-5,13-dienoate
(5Z,9alpha,11alpha,13E,15S)-9,11-epidioxy-15-hydroxy-prosta-5,13-dienoate i.e. prostaglandin H2. Prostaglandin H2 and prostamide H2 are the best substrates. Cys44 is essential for the catalytic activity of prostamide/PGF synthase
687797
Mus musculus
thioredoxin disulfide + (5Z,9alpha,11alpha,13E,15S)-9,11,15-trihydroxyprosta-5,13-dienoate
(5Z,9alpha,11alpha,13E,15S)-9,11,15-trihydroxyprosta-5,13-dienoate i.e. prostaglandin F2alpha
-
-
?
1.11.1.20
thioredoxin + cumene hydroperoxide
-
687797
Mus musculus
thioredoxin disulfide + H2O + 2-phenylpropan-2-ol
-
-
-
?
1.11.1.20
thioredoxin + cumene hydroperoxide
-
687797
Mus musculus
thioredoxin disulfide + ?
-
-
-
?
1.11.1.20
thioredoxin + H2O2
-
687797
Mus musculus
thioredoxin disulfide + 2 H2O
-
-
-
?
1.11.1.20
thioredoxin + H2O2
-
687797
Mus musculus
thioredoxin disulfide + ?
-
-
-
?
1.11.1.20
thioredoxin + prostaglandin H2
prostaglandin H2 is one of the best substrates
687797
Mus musculus
thioredoxin disulfide + prostaglandin F2alpha
-
-
-
?
1.11.1.20
thioredoxin + prostamide H2
prostamide H2 is one of the best substrates
687797
Mus musculus
thioredoxin disulfide + prostamide F2alpha
-
-
-
?
1.11.1.20
thioredoxin + t-butyl hydroperoxide
-
687797
Mus musculus
thioredoxin disulfide + ?
-
-
-
?
1.11.1.20
thioredoxin + tert-butyl hydroperoxide
-
687797
Mus musculus
thioredoxin disulfide + H2O + tert-butanol
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.1.1.188
dimer
2 * 23000, wild-type and mutants, SDS-PAGE
Mus musculus
1.1.1.188
dimer
-
Sus scrofa
1.11.1.20
?
x * 21670, stimulation by ammonium sulfate suggests that the enzyme may be a dimer or oligomer instead of a monomer, calculated from sequence
Mus musculus
1.11.1.20
?
x * 20000, SDS-PAGE
Sus scrofa
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.11.1.20
24
-
assay at
Mus musculus
1.11.1.20
24
-
assay at
Sus scrofa
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.1.1.188
0.2467
-
prostaglandin H2
pH 7.0, 24°C
Mus musculus
1.1.1.188
0.2467
-
prostaglandin H2
-
Sus scrofa
1.11.1.20
0.001
-
thioredoxin
in 0.1 M KPB (pH 7.0), at 24°C
Mus musculus
1.11.1.20
0.09
-
(5Z)-N-(2-hydroxyethyl)-7-[(1R,4S,5R,6R)-6-[(1E,3S)-3-hydroxyoct-1-en-1-yl]-2,3-dioxabicyclo[2.2.1]hept-5-yl]hept-5-enamide
pH 7.0, 24°C, i.e. prostamide H2
Mus musculus
1.11.1.20
0.09
-
prostamide H2
in 0.1 M KPB (pH 7.0), at 24°C
Mus musculus
1.11.1.20
0.09
-
t-butyl hydroperoxide
in 0.1 M KPB (pH 7.0), at 24°C
Mus musculus
1.11.1.20
0.09
-
tert-butyl hydroperoxide
pH 7.0, 24°C
Mus musculus
1.11.1.20
0.1
-
cumene hydroperoxide
in 0.1 M KPB (pH 7.0), at 24°C; pH 7.0, 24°C
Mus musculus
1.11.1.20
0.1
-
H2O2
in 0.1 M KPB (pH 7.0), at 24°C; pH 7.0, 24°C
Mus musculus
1.11.1.20
0.25
-
(5Z,9alpha,11alpha,13E,15S)-9,11-epidioxy-15-hydroxy-prosta-5,13-dienoate
pH 7.0, 24°C, i.e. prostaglandin H2
Mus musculus
1.11.1.20
0.25
-
prostaglandin H2
in 0.1 M KPB (pH 7.0), at 24°C
Mus musculus
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.11.1.20
7
-
; optimal pH for the prostamide F synthase activity
Mus musculus
1.11.1.20
7
-
assay at
Sus scrofa
pH Range (protein specific)
EC Number
pH Minimum
pH Maximum
Commentary
Organism
1.11.1.20
6.5
8
pH 6.5: about 50% of maximal activity, pH 8.0: about 70% of maximal activity
Mus musculus
General Information
EC Number
General Information
Commentary
Organism
1.1.1.188
physiological function
prostamide/PGF synthase may play an important functional role in the central nervous system
Mus musculus
1.1.1.188
physiological function
prostamide/PGF synthase may play an important functional role in the central nervous system. It directly synthesizes prostamide/PGF2alpha from prostamide/PGH2 better than PGF synthase
Sus scrofa
General Information (protein specific)
EC Number
General Information
Commentary
Organism
1.1.1.188
physiological function
prostamide/PGF synthase may play an important functional role in the central nervous system
Mus musculus
1.1.1.188
physiological function
prostamide/PGF synthase may play an important functional role in the central nervous system. It directly synthesizes prostamide/PGF2alpha from prostamide/PGH2 better than PGF synthase
Sus scrofa
KCat/KM [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.1.1.188
35.85
-
prostaglandin H2
pH 7.0, 24°C
Mus musculus
1.11.1.20
0.003
-
H2O2
in 0.1 M KPB (pH 7.0), at 24°C
Mus musculus
1.11.1.20
0.22
-
t-butyl hydroperoxide
in 0.1 M KPB (pH 7.0), at 24°C
Mus musculus
1.11.1.20
3.9
-
cumene hydroperoxide
in 0.1 M KPB (pH 7.0), at 24°C
Mus musculus
1.11.1.20
11.88
-
prostamide H2
in 0.1 M KPB (pH 7.0), at 24°C
Mus musculus
1.11.1.20
35.85
-
prostaglandin H2
in 0.1 M KPB (pH 7.0), at 24°C
Mus musculus
KCat/KM [mM/s] (protein specific)
EC Number
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.1.1.188
35.85
-
prostaglandin H2
pH 7.0, 24°C
Mus musculus
1.11.1.20
0.003
-
H2O2
in 0.1 M KPB (pH 7.0), at 24°C
Mus musculus
1.11.1.20
0.22
-
t-butyl hydroperoxide
in 0.1 M KPB (pH 7.0), at 24°C
Mus musculus
1.11.1.20
3.9
-
cumene hydroperoxide
in 0.1 M KPB (pH 7.0), at 24°C
Mus musculus
1.11.1.20
11.88
-
prostamide H2
in 0.1 M KPB (pH 7.0), at 24°C
Mus musculus
1.11.1.20
35.85
-
prostaglandin H2
in 0.1 M KPB (pH 7.0), at 24°C
Mus musculus