Biochemical characterization of mouse microsomal prostaglandin E synthase-1 and its colocalization with cyclooxygenase-2 in peritoneal macrophages

Arch Biochem Biophys. 2002 Jan 15;397(2):336-41. doi: 10.1006/abbi.2001.2614.


We cloned the cDNA for mouse microsomal prostaglandin (PG) E synthase-1 (mPGES-1) and expressed the recombinant enzyme in Escherichia coli. The membrane fraction containing recombinant mPGES-1 catalyzed the isomerization of PGH2 to PGE2 in the presence of GSH with K(m) values of 130 microM for PGH2 and 37 microM for GSH, a turnover number of 600 min(-1), and a k(cat)/K(m) ratio of 4.6 min(-1) microM(-1). Recombinant mPGES-1 was purified and used to generate a polyclonal antibody highly specific for mPGES-1. The antibody showed a single band on Western blotting of microsomal fractions from lipopolysaccharide-treated mouse peritoneal macrophages. Northern and Western blotting analyses revealed that mPGES-1 was induced together with cyclooxygenase-2 in mouse macrophages after treatment of the cells with lipopolysaccharide. Confocal immunofluorescence microscopy revealed that both mPGES-1 and cyclooxygenase-2 were colocalized in the lipopolysaccharide-treated macrophages. Taken together, these results demonstrate that mPGES-1 is an efficient downstream enzyme for the production of PGE2 in the activated macrophages treated by lipopolysaccharide.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cloning, Molecular
  • Cyclooxygenase 2
  • Guinea Pigs
  • Intramolecular Oxidoreductases / genetics
  • Intramolecular Oxidoreductases / isolation & purification*
  • Isoenzymes / isolation & purification*
  • Kinetics
  • Lipopolysaccharides / pharmacology
  • Macrophages, Peritoneal / drug effects
  • Macrophages, Peritoneal / enzymology*
  • Mice
  • Mice, Inbred C57BL
  • Microsomes / enzymology*
  • Molecular Sequence Data
  • Prostaglandin-E Synthases
  • Prostaglandin-Endoperoxide Synthases / isolation & purification*


  • Isoenzymes
  • Lipopolysaccharides
  • Cyclooxygenase 2
  • Prostaglandin-Endoperoxide Synthases
  • Intramolecular Oxidoreductases
  • Prostaglandin-E Synthases
  • Ptges protein, mouse

Associated data

  • GENBANK/AB035323