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Expression, Purification and Characterization of Recombinant Human Microsomal PGE2 Synthase-1

  • Marc Ouellet
  • Ally Pen
  • Po-Hien Ear
  • Jean-Pierre Falgueyret
  • Tammy G. LeRiche
  • Joseph A. Mancini
  • Denis Riendeau
  • M. David Percival
Chapter
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 525)

Abstract

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 [1], whereas a constitutively and widely expressed cytosolic PGES (cPGES or p23) is believed to couple with Cox-1 [2]. 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 [3]. 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 [4]. 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.

Keywords

Lipoic Acid Octyl Glucoside Wide Tissue Distribution Hydroxyapatite Column Cytosolic Glutathione 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer Science+Business Media New York 2003

Authors and Affiliations

  • Marc Ouellet
    • 1
  • Ally Pen
    • 1
  • Po-Hien Ear
    • 1
  • Jean-Pierre Falgueyret
    • 1
  • Tammy G. LeRiche
    • 1
  • Joseph A. Mancini
    • 1
  • Denis Riendeau
    • 1
  • M. David Percival
    • 1
  1. 1.Department of Biochemistry and Molecular BiologyMerck Frosst CanadaKirklandCanada

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