Purification and characterization of membrane-bound prostaglandin E synthase from bovine heart

Biochim Biophys Acta. 1999 Aug 18;1439(3):406-14. doi: 10.1016/s1388-1981(99)00084-0.


Prostaglandin (PG) E synthase was solubilized with 6 mM sodium deoxycholate from the microsomal fraction of bovine hearts. The enzyme was purified by about 800-fold to apparent homogeneity. The specific activity of the purified enzyme was about 830 mU/mg of protein, and the K(m) value for PGH(2) was 24 microM. The molecular weight of the enzyme was about 31000 on SDS-polyacrylamide gel electrophoresis and was about 60000 by gel filtration. The enzyme was separated from glutathione (GSH) S-transferase by DEAE-Toyopearl column chromatography, and did not exhibit any GSH S-transferase activity towards four different substrates. The purified enzyme was active in the absence of GSH, but it was activated by various SH-reducing reagents including dithiothreitol, GSH, or beta-mercaptoethanol. This is the first reported purification of membrane-bound PGE synthase to apparent homogeneity.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cattle
  • Deoxycholic Acid
  • Detergents
  • Dithiothreitol / pharmacology
  • Enzyme Activation / drug effects
  • Glutathione / pharmacology
  • Glutathione Transferase / analysis
  • Hydrogen-Ion Concentration
  • Intracellular Membranes / enzymology*
  • Intramolecular Oxidoreductases / chemistry
  • Intramolecular Oxidoreductases / isolation & purification*
  • Intramolecular Oxidoreductases / metabolism
  • Male
  • Mercaptoethanol / pharmacology
  • Microsomes / enzymology
  • Myocardium / enzymology*
  • Prostaglandin-E Synthases
  • Sheep
  • Solubility


  • Detergents
  • Deoxycholic Acid
  • Mercaptoethanol
  • Glutathione Transferase
  • Intramolecular Oxidoreductases
  • Prostaglandin-E Synthases
  • Glutathione
  • Dithiothreitol