BRENDA - Enzyme Database show

New insights into the catalytic mechanism of Bombyx mori prostaglandin E synthase gained from structure-function analysis

Yamamoto, K.; Suzuki, M.; Higashiura, A.; Aritake, K.; Urade, Y.; Uodome, N.; Hossain, T.; Nakagawa, A.; Biochem. Biophys. Res. Commun. 440, 762-767 (2013)

Data extracted from this reference:

Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
5.3.99.3
glutathione
the N-terminal domain contains the GSH-binding site (G-site)
Bombyx mori
Cloned(Commentary)
EC Number
Commentary
Organism
5.3.99.3
recombinant expression
Bombyx mori
Crystallization (Commentary)
EC Number
Crystallization
Organism
5.3.99.3
purified enzyme in complex with glutathione sulfonic acid, sitting drop vapor diffusion method, mixing of 10 mg/ml protein in 20 mM Tris-HCl , pH 8.5, and 0.2 M NaCl with 0.1 M HEPES, pH 7.5, 30% PEG 400 w/v, and 20% 1,2-propanediol v/v, in a 1:1 or 1:2 ratio, X-ray diffraction structure determination and analysis at 1.37 A resolution, molecular replacement
Bombyx mori
Engineering
EC Number
Amino acid exchange
Commentary
Organism
5.3.99.3
D97A
site-directed mutagenesis, isomerase inactive mutant
Bombyx mori
5.3.99.3
N96A
site-directed mutagenesis, the mutant shows reduced isomerase activity compared to the wild-type enzyme
Bombyx mori
5.3.99.3
R99A
site-directed mutagenesis, isomerase inactive mutant
Bombyx mori
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
5.3.99.3
(5Z,13E,15S)-9alpha,11alpha-epidioxy-15-hydroxyprosta-5,13-dienoate
Bombyx mori
-
(5Z,13E,15S)-11alpha,15-dihydroxy-9-oxoprosta-5,13-dienoate
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
5.3.99.3
Bombyx mori
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
5.3.99.3
recombinant enzyme by ammonium sulfate fractionation, ion exchange chromatography, and gel filtration
Bombyx mori
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
5.3.99.3
(5Z,13E,15S)-9alpha,11alpha-epidioxy-15-hydroxyprosta-5,13-dienoate
-
726884
Bombyx mori
(5Z,13E,15S)-11alpha,15-dihydroxy-9-oxoprosta-5,13-dienoate
-
-
-
?
5.3.99.3
additional information
the enzyme also shows glutathione transferase activity with 1-chloro-2,4-dinitrobenzene
726884
Bombyx mori
?
-
-
-
-
Subunits
EC Number
Subunits
Commentary
Organism
5.3.99.3
More
the N-terminal domain contains the GSH-binding site (G-site), and the C-terminal domain contains the binding site for hydrophobic substrate and PGH2
Bombyx mori
Temperature Optimum [C]
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
5.3.99.3
25
-
assay at
Bombyx mori
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
5.3.99.3
8
-
assay at
Bombyx mori
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
5.3.99.3
glutathione
the N-terminal domain contains the GSH-binding site (G-site)
Bombyx mori
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
5.3.99.3
recombinant expression
Bombyx mori
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
5.3.99.3
purified enzyme in complex with glutathione sulfonic acid, sitting drop vapor diffusion method, mixing of 10 mg/ml protein in 20 mM Tris-HCl , pH 8.5, and 0.2 M NaCl with 0.1 M HEPES, pH 7.5, 30% PEG 400 w/v, and 20% 1,2-propanediol v/v, in a 1:1 or 1:2 ratio, X-ray diffraction structure determination and analysis at 1.37 A resolution, molecular replacement
Bombyx mori
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
5.3.99.3
D97A
site-directed mutagenesis, isomerase inactive mutant
Bombyx mori
5.3.99.3
N96A
site-directed mutagenesis, the mutant shows reduced isomerase activity compared to the wild-type enzyme
Bombyx mori
5.3.99.3
R99A
site-directed mutagenesis, isomerase inactive mutant
Bombyx mori
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
5.3.99.3
(5Z,13E,15S)-9alpha,11alpha-epidioxy-15-hydroxyprosta-5,13-dienoate
Bombyx mori
-
(5Z,13E,15S)-11alpha,15-dihydroxy-9-oxoprosta-5,13-dienoate
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
5.3.99.3
recombinant enzyme by ammonium sulfate fractionation, ion exchange chromatography, and gel filtration
Bombyx mori
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
5.3.99.3
(5Z,13E,15S)-9alpha,11alpha-epidioxy-15-hydroxyprosta-5,13-dienoate
-
726884
Bombyx mori
(5Z,13E,15S)-11alpha,15-dihydroxy-9-oxoprosta-5,13-dienoate
-
-
-
?
5.3.99.3
additional information
the enzyme also shows glutathione transferase activity with 1-chloro-2,4-dinitrobenzene
726884
Bombyx mori
?
-
-
-
-
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
5.3.99.3
More
the N-terminal domain contains the GSH-binding site (G-site), and the C-terminal domain contains the binding site for hydrophobic substrate and PGH2
Bombyx mori
Temperature Optimum [C] (protein specific)
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
5.3.99.3
25
-
assay at
Bombyx mori
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
5.3.99.3
8
-
assay at
Bombyx mori
General Information
EC Number
General Information
Commentary
Organism
5.3.99.3
evolution
the enzyme belongs to Sigma class glutathione transferase
Bombyx mori
5.3.99.3
additional information
the electron-sharing network of the Bombyx mori enzyme includes Asn95, Asp96, and Arg98, the residues contribute to catalytic activity. The C-terminal domain contains the binding site for hydrophobic substrate and PGH2, active site and substrate binding site structures, overview
Bombyx mori
General Information (protein specific)
EC Number
General Information
Commentary
Organism
5.3.99.3
evolution
the enzyme belongs to Sigma class glutathione transferase
Bombyx mori
5.3.99.3
additional information
the electron-sharing network of the Bombyx mori enzyme includes Asn95, Asp96, and Arg98, the residues contribute to catalytic activity. The C-terminal domain contains the binding site for hydrophobic substrate and PGH2, active site and substrate binding site structures, overview
Bombyx mori