Expression, Purification and Characterization of Recombinant Human Microsomal PGE2 Synthase-1
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Prostaglandin E synthases (PGES) comprise a number of structurally different enzymes that convert the product of the cyclooxygenase enzymes, PGH2, into the physiologically important PGE2. An inducible microsomal PGES (mPGES-1) couples preferentially with COX-2 , whereas a constitutively and widely expressed cytosolic PGES (cPGES or p23) is believed to couple with Cox-1 . Both of these enzyme activities are dependent on GSH. Most recently, a second, membrane-associated PGES (mPGES-2) that has a wide tissue distribution and is activated by a number of thio1-containing reagents was identified and purified . Two cytosolic glutathione transferases (μ2 and μ3) also have some PGES activity. As the expression of mPGES-1 is strongly induced by pro-inflammatory stimuli and is down-regulated by glucocorticoids, mPGES-1 is a potential drug target for anti-inflammatory therapy. A solubilization and partial purification of a microsomal PGES activity from bovine vesicular glands has been reported . It appears that this enzyme is the same as mPGES-1. Here we report the baculovirus over-expression of recombinant human mPGES-1 in Sf9 insect cells, its purification to apparent homogeneity and the characterization of its enzymatic properties.
KeywordsLipoic Acid Octyl Glucoside Wide Tissue Distribution Hydroxyapatite Column Cytosolic Glutathione
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