Detection and isolation of an endoperoxide intermediate in prostaglandin biosynthesis

Proc Natl Acad Sci U S A. 1973 Mar;70(3):899-903. doi: 10.1073/pnas.70.3.899.

Abstract

An earlier proposed endoperoxide intermediate in the biosynthesis of prostaglandins was detected in short-time incubations of arachidonic acid with the microsomal fraction of homogenates of sheep vesicular glands. Conversion of the endoperoxide into prostaglandin E(2) was stimulated by reduced glutathione but suppressed by p-mercuribenzoate and N-ethylmaleimide. The methyl ester of an unknown compound was isolated by solvent extraction and thin-layer chromatography after short-time incubation of arachidonic acid with the microsomal fraction and p-mercuribenzoate. This derivative was identical to the methylester of the endoperoxide, as shown by its conversion into the methyl esters of 11-dehydroprostaglandin F(2alpha) and prostaglandin E(2) by spontaneous rearrangement and its conversion into the methyl ester of prostaglandin F(2alpha) by mild chemical reduction. The smooth muscle-stimulating activity of the endoperoxide ester on the isolated rabbit aortas trip was 4- to 8-times higher than that of the methyl ester of prostaglandin E(2).

MeSH terms

  • Animals
  • Aorta / drug effects
  • Arachidonic Acids / metabolism
  • Biological Assay
  • Carbon Isotopes
  • Chromatography, Thin Layer
  • Deuterium
  • In Vitro Techniques
  • Kinetics
  • Male
  • Microsomes / metabolism
  • Peroxides / isolation & purification
  • Peroxides / metabolism*
  • Prostaglandins / analysis
  • Prostaglandins / biosynthesis*
  • Prostaglandins / pharmacology
  • Rabbits
  • Seminal Vesicles / cytology
  • Seminal Vesicles / metabolism
  • Sheep

Substances

  • Arachidonic Acids
  • Carbon Isotopes
  • Peroxides
  • Prostaglandins
  • Deuterium