BRENDA - Enzyme Database show

Crystallization and preliminary X-ray crystallographic studies of Trypanosoma brucei prostaglandin F2 alpha synthase

Okano, Y.; Inoue, T.; Kubata, B.K.; Kabututu, Z.; Urade, Y.; Matsumura, H.; Kai, Y.; J. Biochem. 132, 859-861 (2002)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
1.1.1.188
expression in Escherichia coli strain BL21(DE3) as GST-fusion protein
Trypanosoma brucei
Crystallization (Commentary)
EC Number
Crystallization
Organism
1.1.1.188
10 mg/ml purified recombinant enzyme from 0.1 M HEPES/NaOH, pH 7.5, 2.0 M ammonium sulfate, 5% w/v PEG 400, and 0.5 M NADPH by hanging drop vapour diffusion method, 20°C, 3 weeks, X-ray diffraction structure determination at room temperature and analysis at 2.6 A resolution
Trypanosoma brucei
Localization
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
1.1.1.188
cytosol
-
Trypanosoma brucei
5829
-
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.1.1.188
prostaglandin H2 + NADPH
Trypanosoma brucei
-
prostaglandin F2alpha + NADP+
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.1.1.188
Trypanosoma brucei
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
1.1.1.188
recombinant enzyme from Escherichia coli by GSH affinity chromatography, cleavage of the fusion protein glutathione S-tranferase by thrombin, and DEAE ion exchange chromatography
Trypanosoma brucei
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.1.1.188
9,10-phenanthrenequinone + NADPH
-
655915
Trypanosoma brucei
?
-
-
-
?
1.1.1.188
additional information
enzyme does not reduce prostaglandins D2 and E2
655915
Trypanosoma brucei
?
-
-
-
-
1.1.1.188
prostaglandin H2 + NADPH
-
655915
Trypanosoma brucei
prostaglandin F2alpha + NADP+
-
-
-
?
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.1.1.188
37
-
assay at
Trypanosoma brucei
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.1.1.188
7
-
assay at
Trypanosoma brucei
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.188
NADPH
dependent on
Trypanosoma brucei
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.188
expression in Escherichia coli strain BL21(DE3) as GST-fusion protein
Trypanosoma brucei
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.188
NADPH
dependent on
Trypanosoma brucei
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
1.1.1.188
10 mg/ml purified recombinant enzyme from 0.1 M HEPES/NaOH, pH 7.5, 2.0 M ammonium sulfate, 5% w/v PEG 400, and 0.5 M NADPH by hanging drop vapour diffusion method, 20°C, 3 weeks, X-ray diffraction structure determination at room temperature and analysis at 2.6 A resolution
Trypanosoma brucei
Localization (protein specific)
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
1.1.1.188
cytosol
-
Trypanosoma brucei
5829
-
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.1.1.188
prostaglandin H2 + NADPH
Trypanosoma brucei
-
prostaglandin F2alpha + NADP+
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.188
recombinant enzyme from Escherichia coli by GSH affinity chromatography, cleavage of the fusion protein glutathione S-tranferase by thrombin, and DEAE ion exchange chromatography
Trypanosoma brucei
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
9,10-phenanthrenequinone + NADPH
-
655915
Trypanosoma brucei
?
-
-
-
?
1.1.1.188
additional information
enzyme does not reduce prostaglandins D2 and E2
655915
Trypanosoma brucei
?
-
-
-
-
1.1.1.188
prostaglandin H2 + NADPH
-
655915
Trypanosoma brucei
prostaglandin F2alpha + NADP+
-
-
-
?
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.1.1.188
37
-
assay at
Trypanosoma brucei
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.1.1.188
7
-
assay at
Trypanosoma brucei