Cloned (Comment) | Organism |
---|---|
gene P100/11E, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3) | Leishmania major |
gene Tc00.1047053511287.49, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3) | Trypanosoma cruzi |
Crystallization (Comment) | Organism |
---|---|
purified recombinant apoenzyme or enzyme with bound cofactor NADP+, mixing of 20 mg/ml protein in 25 mM Tris pH 8.0, 200 mM NaCl, 1% v/v glycerol, 1 mM TCEP with reservoir solution. For the apoenzyme, the reservoir solution contains 0.1 M MES-imidazole, pH 6.5, 10% PEG 20 000, 20% PEG 550 MME, 0.02 M glutamic acid, glycine, serine, alanine, and lysine, and for the cofactor complexed enzyme, it contains 0.2 M ammonium acetate, 0.1 M Bis-Tris pH 5.5, 25% PEG 3350, cryoprotectant is 15% ethylene glycol, 16°C, 2-4 weeks, X-ray diffraction structure determination and analysis at 1.25 A and 2.6 A resolution, respectively, molecular replacement | Trypanosoma cruzi |
purified recombinant apoenzyme or enzyme with bound cofactor NADPH, mixing of 20 mg/ml protein in 25 mM Tris, pH 8.0, 200 mM NaCl, 1% v/v glycerol, 1 mM TCEP with the reservoir solution containing 0.1 M sodium citrate pH 5.50, 20% PEG 3000 200 mM magnesium chloride, 100 mM Tris, pH 7.0, 10% PEG 8000 for apo- and complexed enzyme, 16°C, 2-4 weeks, X-ray diffraction structure determination and analysis at 1.8 A and 1.6 A resolution, respectively, molecular replacement | Leishmania major |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(5Z,13E)-(15S)-9alpha,11alpha,15-trihydroxyprosta-5,13-dienoate + NADP+ | Leishmania major | - |
(5Z,13E)-(15S)-9alpha,15-dihydroxy-11-oxoprosta-5,13-dienoate + NADPH + H+ | - |
? | |
(5Z,13E)-(15S)-9alpha,11alpha,15-trihydroxyprosta-5,13-dienoate + NADP+ | Trypanosoma cruzi | - |
(5Z,13E)-(15S)-9alpha,15-dihydroxy-11-oxoprosta-5,13-dienoate + NADPH + H+ | - |
? | |
(5Z,13E)-(15S)-9alpha,11alpha,15-trihydroxyprosta-5,13-dienoate + NADP+ | Trypanosoma cruzi CL Brener | - |
(5Z,13E)-(15S)-9alpha,15-dihydroxy-11-oxoprosta-5,13-dienoate + NADPH + H+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Leishmania major | P22045 | - |
- |
Trypanosoma cruzi | Q4DJ07 | - |
- |
Trypanosoma cruzi CL Brener | Q4DJ07 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant N-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration followed by a polyether sulfon concentration | Leishmania major |
recombinant N-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration followed by a polyether sulfon concentration | Trypanosoma cruzi |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(5Z,13E)-(15S)-9alpha,11alpha,15-trihydroxyprosta-5,13-dienoate + NADP+ | - |
Leishmania major | (5Z,13E)-(15S)-9alpha,15-dihydroxy-11-oxoprosta-5,13-dienoate + NADPH + H+ | - |
? | |
(5Z,13E)-(15S)-9alpha,11alpha,15-trihydroxyprosta-5,13-dienoate + NADP+ | - |
Trypanosoma cruzi | (5Z,13E)-(15S)-9alpha,15-dihydroxy-11-oxoprosta-5,13-dienoate + NADPH + H+ | - |
? | |
(5Z,13E)-(15S)-9alpha,11alpha,15-trihydroxyprosta-5,13-dienoate + NADP+ | - |
Trypanosoma cruzi CL Brener | (5Z,13E)-(15S)-9alpha,15-dihydroxy-11-oxoprosta-5,13-dienoate + NADPH + H+ | - |
? |
Synonyms | Comment | Organism |
---|---|---|
LemaA.00019.a.B1 | - |
Leishmania major |
P100/11E | - |
Leishmania major |
PGF | - |
Leishmania major |
PGF | - |
Trypanosoma cruzi |
prostaglandin F synthase | - |
Leishmania major |
prostaglandin F synthase | - |
Trypanosoma cruzi |
Tc00.1047053511287.49 | - |
Trypanosoma cruzi |
TrcrA.00019.a.B1 | - |
Trypanosoma cruzi |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADP+ | enzyme PGF is specific for NADP+. Structures of PGF with and without NADP+ and structural differences between the bound and the unbound state of the enzyme with regard to its cofactor | Leishmania major | |
NADP+ | enzyme PGF is specific for NADP+. Structures of PGF with and without NADP+ and structural differences between the bound and the unbound state of the enzyme with regard to its cofactor | Trypanosoma cruzi |
General Information | Comment | Organism |
---|---|---|
metabolism | enzyme PGF is involved in essential lipid metabolism pathways | Leishmania major |
metabolism | enzyme PGF is involved in essential lipid metabolism pathways | Trypanosoma cruzi |
additional information | a loop with residues 188-196 in TrcrA.00019.a.B1 shows significant movement between the apoenzyme and holoenzyme. The Trypanosoma cruzi loop becomes ordered and a key interaction with one of the phosphates of NADP at Ser193 is likely to stabilize this region | Trypanosoma cruzi |
additional information | a loop with residues 201-205 (disordered) in LemaA.00019.a.B1 shows significant movement between the apoenzyme and holoenzyme. The Leishmania major loop becomes ordered and a key interaction with one of the phosphates of NADP at Gln202 is likely to stabilize this region | Leishmania major |