prostaglandin-I synthase / prostaglandin-I synthase activity / Eicosanoids / Sterols are 12-hydroxylated by CYP8B1 / icosanoid metabolic process / hydroperoxy icosatetraenoate dehydratase / hydroperoxy icosatetraenoate dehydratase activity / positive regulation of peroxisome proliferator activated receptor signaling pathway / NAD biosynthesis via nicotinamide riboside salvage pathway / Synthesis of bile acids and bile salts via 24-hydroxycholesterol ...prostaglandin-I synthase / prostaglandin-I synthase activity / Eicosanoids / Sterols are 12-hydroxylated by CYP8B1 / icosanoid metabolic process / hydroperoxy icosatetraenoate dehydratase / hydroperoxy icosatetraenoate dehydratase activity / positive regulation of peroxisome proliferator activated receptor signaling pathway / NAD biosynthesis via nicotinamide riboside salvage pathway / Synthesis of bile acids and bile salts via 24-hydroxycholesterol / Synthesis of Prostaglandins (PG) and Thromboxanes (TX) / negative regulation of nitric oxide biosynthetic process / Nicotinamide salvaging / cyclooxygenase pathway / cellular response to interleukin-6 / oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen / prostaglandin biosynthetic process / negative regulation of NF-kappaB transcription factor activity / positive regulation of execution phase of apoptosis / cellular response to interleukin-1 / Synthesis of bile acids and bile salts via 27-hydroxycholesterol / Synthesis of bile acids and bile salts via 7alpha-hydroxycholesterol / monooxygenase activity / caveola / apoptotic signaling pathway / negative regulation of inflammatory response / positive regulation of angiogenesis / cellular response to hypoxia / iron ion binding / heme binding / endoplasmic reticulum membrane / endoplasmic reticulum / extracellular space / nucleus Similarity search - Function
Mass: 18.015 Da / Num. of mol.: 765 / Source method: isolated from a natural source / Formula: H2O
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Experimental details
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Experiment
Experiment
Method: X-RAY DIFFRACTION / Number of used crystals: 1
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Sample preparation
Crystal
Density Matthews: 2.35 Å3/Da / Density % sol: 47.7 %
Crystal grow
Temperature: 277 K / Method: vapor diffusion, hanging drop / pH: 8 Details: Crystallization of native human prostacyclin synthase (hPGIS). The native recombinant PGIS (25 mg/ml in the gel filtration buffer containing 20 mM Tris, pH 8.0 and 150 mM NaCl) was found to ...Details: Crystallization of native human prostacyclin synthase (hPGIS). The native recombinant PGIS (25 mg/ml in the gel filtration buffer containing 20 mM Tris, pH 8.0 and 150 mM NaCl) was found to crystallize spontaneously into needle clusters in the Eppendorf tube when stored at 4 C for about a week. A combination of microseeding technique and the hanging-drop vapor-diffusion method was then used to improve crystal quality. Specifically, seed stock was prepared by crushing these initial PGIS needle clusters with the Seed Bead Kit (Hampton Research). Following four 10-fold serial dilutions of the seed stock using the gel filtration buffer, 1 microliter of diluted seeds was added into a 10 microliter drop of freshly concentrated PGIS sample, and equilibrated at 4 C against 450 microliter of the gel filtration buffer. Crystallization of hPGIS-minoxidil complex: The inhibitor-enzyme complex was prepared by adding minoxidil (final concentration 0.1 mM) to a 20 mg/ml hPGIS stock solution. Crystals of the complex were then obtained by co-crystallization, with the technique described above using ligand-free hPGIS microcrystals as seeds., VAPOR DIFFUSION, HANGING DROP, temperature 277K
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