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Literature summary extracted from

  • Moriuchi, H.; Koda, N.; Okuda-Ashitaka, E.; Daiyasu, H.; Ogasawara, K.; Toh, H.; Ito, S.; Woodward, D.F.; Watanabe, K.
    Molecular characterization of a novel type of prostamide/prostaglandin F synthase, belonging to the thioredoxin-like superfamily (2008), J. Biol. Chem., 283, 792-801.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment 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 Cloned (Comment) 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 Mus musculus
1.11.1.20 expressed in Escherichia coli BL21(DE3) cells Mus musculus

Protein Variants

EC Number Protein Variants Comment 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 Mus musculus
1.11.1.20 C44S 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 Mus musculus
1.11.1.20 C44S/C47S 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 Mus musculus
1.11.1.20 C47S retains 63% of wild-type activity Mus musculus

Inhibitors

EC Number Inhibitors Comment 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 Comment 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 pH 7.0, 24°C Mus musculus
1.11.1.20 0.026
-
 cumene 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 0.4
-
 t-butyl hydroperoxide in 0.1 M KPB (pH 7.0), at 24°C Mus musculus
1.11.1.20 1.6
-
 H2O2 pH 7.0, 24°C Mus musculus

Localization

EC Number Localization Comment 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 Comment 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] Comment 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 UniProt Comment 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 Purification (Comment) 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
-
 Mus musculus
1.11.1.20 Ni2+-agarose column chromatography and Q Sepharose column chromatography Mus musculus

Source Tissue

EC Number Source Tissue Comment Organism Textmining
1.1.1.188 adrenal gland
-
 Mus musculus
-
1.1.1.188 brain highest activity Sus scrofa
-
1.1.1.188 brain high activity Mus musculus
-
1.1.1.188 cecum low activity Mus musculus
-
1.1.1.188 eye
-
 Sus scrofa
-
1.1.1.188 eye low activity Mus musculus
-
1.1.1.188 heart
-
 Sus scrofa
-
1.1.1.188 heart high activity Mus musculus
-
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
-
 Sus scrofa
-
1.1.1.188 lung low activity Mus musculus
-
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
-
 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
-
 Sus scrofa
-
1.11.1.20 brain 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 highest activity in the spinal cord and brain Mus musculus
-
1.11.1.20 cecum lowest activity in the cecum Mus musculus
-
1.11.1.20 epididymis
-
 Mus musculus
-
1.11.1.20 heart
-
 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
-
 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 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 spinal cord highest activity in the spinal cord and brain Mus musculus
-
1.11.1.20 stomach
-
 Mus musculus
-
1.11.1.20 testis
-
 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
-
 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
-
 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
-
 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] Comment 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 Mus musculus
1.11.1.20 0.25
-
 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 Mus musculus
1.11.1.20 0.28
-
 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 Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.188 additional information no activity with prostaglandin D2 and prostaglandin E2 Mus musculus ?
-
 ?
1.1.1.188 prostaglandin H2 + NADPH + H+
-
 Mus musculus prostaglandin F2alpha + NADP+
-
 ?
1.1.1.188 prostaglandin H2 + NADPH + H+
-
 Sus scrofa prostaglandin F2alpha + NADP+
-
 ?
1.11.1.20 additional information prostaglandin D2 and prostaglandin E2 are not reduced 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 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 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 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 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 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 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
-
 Mus musculus thioredoxin disulfide + H2O + 2-phenylpropan-2-ol
-
 ?
1.11.1.20 thioredoxin + cumene hydroperoxide
-
 Mus musculus thioredoxin disulfide + ?
-
 ?
1.11.1.20 thioredoxin + H2O2
-
 Mus musculus thioredoxin disulfide + 2 H2O
-
 ?
1.11.1.20 thioredoxin + H2O2
-
 Mus musculus thioredoxin disulfide + ?
-
 ?
1.11.1.20 thioredoxin + prostaglandin H2 prostaglandin H2 is one of the best substrates Mus musculus thioredoxin disulfide + prostaglandin F2alpha
-
 ?
1.11.1.20 thioredoxin + prostamide H2 prostamide H2 is one of the best substrates Mus musculus thioredoxin disulfide + prostamide F2alpha
-
 ?
1.11.1.20 thioredoxin + t-butyl hydroperoxide
-
 Mus musculus thioredoxin disulfide + ?
-
 ?
1.11.1.20 thioredoxin + tert-butyl hydroperoxide
-
 Mus musculus thioredoxin disulfide + H2O + tert-butanol
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.1.188 dimer
-
 Sus scrofa
1.1.1.188 dimer 2 * 23000, wild-type and mutants, SDS-PAGE Mus musculus
1.11.1.20 ? x * 20000, SDS-PAGE 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

Synonyms

EC Number Synonyms Comment Organism
1.1.1.188 prostamide/PGF synthase
-
 Mus musculus
1.1.1.188 prostamide/PGF synthase
-
 Sus scrofa
1.1.1.188 prostamide/prostaglandin F synthase
-
 Mus musculus
1.1.1.188 prostamide/prostaglandin F synthase
-
 Sus scrofa
1.11.1.20 PGF prostamide F synthase
-
 Mus musculus
1.11.1.20 prostaglandin F ethanolamide synthase
-
 Mus musculus
1.11.1.20 prostamide F synthase
-
 Sus scrofa
1.11.1.20 prostamide F synthase
-
 Mus musculus
1.11.1.20 prostamide/PGF synthase
-
 Sus scrofa
1.11.1.20 prostamide/PGF synthase
-
 Mus musculus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.11.1.20 24
-
 assay at Sus scrofa
1.11.1.20 24
-
 assay at Mus musculus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.188 0.2467
-
 prostaglandin H2
-
 Sus scrofa
1.1.1.188 0.2467
-
 prostaglandin H2 pH 7.0, 24°C Mus musculus
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
-
 tert-butyl hydroperoxide pH 7.0, 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
-
 t-butyl hydroperoxide in 0.1 M KPB (pH 7.0), at 24°C 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 0.1
-
 H2O2 pH 7.0, 24°C Mus musculus
1.11.1.20 0.1
-
 cumene hydroperoxide 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 Mus musculus
1.11.1.20 0.1
-
 cumene hydroperoxide in 0.1 M KPB (pH 7.0), at 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 Comment Organism
1.11.1.20 7
-
-
 Mus musculus
1.11.1.20 7
-
 assay at Sus scrofa
1.11.1.20 7
-
 optimal pH for the prostamide F synthase activity Mus musculus

pH Range

EC Number pH Minimum pH Maximum Comment 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 Comment Organism Structure
1.1.1.188 NADPH
-
 Mus musculus
1.1.1.188 NADPH
-
 Sus scrofa
1.11.1.20 thioredoxin
-
 Mus musculus

General Information

EC Number General Information Comment 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 Comment 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