Liver microsomal expoxide hydrase. Solubilization, purification, and characterization

J Biol Chem. 1975 Oct 25;250(20):8283-8.

Abstract

Epoxide hydrase was solubilized from liver microsomes of phenobarbital-treated rats by treatment with cholate and purified to apparent homogeneity by ammonium sulfate fractionation and column chromatography in the presence of the nonionic detergent Emulgen 911 on DEAE-cellulose and hydroxylapatite. The purified enzyme preparation had a single major band with a molecular weight of 53,000 to 54,000 on polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Other studies indicated that in the absence of sodium dodecyl sulfate, purified epoxide hydrase exists as high molecular weight aggregates. The preparation was essentially free of heme and flavin, but still contained small amounts of lipids and Emulgen 911.

MeSH terms

  • Amino Acids / analysis
  • Animals
  • Chelating Agents / pharmacology
  • Epoxide Hydrolases / isolation & purification*
  • Epoxide Hydrolases / metabolism
  • Hydro-Lyases / isolation & purification*
  • Hydrogen-Ion Concentration
  • Kinetics
  • Male
  • Microsomes, Liver / enzymology*
  • Molecular Weight
  • Rats
  • Solubility
  • Spectrophotometry
  • Spectrophotometry, Ultraviolet
  • Sulfhydryl Reagents / pharmacology

Substances

  • Amino Acids
  • Chelating Agents
  • Sulfhydryl Reagents
  • Epoxide Hydrolases
  • Hydro-Lyases