Co-purification of microsomal epoxide hydrolase with the warfarin-sensitive vitamin K1 oxide reductase of the vitamin K cycle

Biochem Pharmacol. 1998 Jan 15;55(2):169-75. doi: 10.1016/s0006-2952(97)00431-0.

Abstract

Vitamin K1 oxide reductase activity has been partially purified from rat liver microsomes. A three-step procedure produced a preparation in which warfarin-sensitive vitamin K1 oxide reductase activity was 118-fold enriched over the activity in intact rat liver microsomes. A major component of the multi-protein mixture was identified as a 50 kDa protein that strongly cross-reacts with antiserum prepared against homogeneous rat liver microsomal epoxide hydrolase. The reductase preparation also had a high level or epoxide hydrolase activity against two xenobiotic epoxide substrates. The K(m) values for hydrolysis by the reductase preparation were similar to those for homogeneous microsomal epoxide hydrolase itself, and the specific hydrolase activities of the reductase preparation were 25-35% of the specific activities measured for the homogeneous hydrolase preparation. Antibodies prepared against homogeneous microsomal epoxide hydrolase inhibited up to 80% of reductase activity of the reductase preparation. Homogeneous microsomal epoxide hydrolase had no vitamin K1 oxide reductase activity. This evidence suggests that microsomal epoxide hydrolase, or a protein that is very similar to it, is a major functional component of a multi-protein complex that is responsible for vitamin K1 oxide reduction in rat liver microsomes.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Chromatography
  • Chromatography, Gel
  • Cross Reactions
  • Durapatite
  • Electrophoresis, Polyacrylamide Gel
  • Epoxide Hydrolases / isolation & purification*
  • Epoxide Hydrolases / metabolism
  • Immune Sera
  • Kinetics
  • Microsomes, Liver / enzymology*
  • Mixed Function Oxygenases / isolation & purification*
  • Mixed Function Oxygenases / metabolism
  • Molecular Weight
  • Rats
  • Vitamin K Epoxide Reductases
  • Warfarin / pharmacology*

Substances

  • Immune Sera
  • Warfarin
  • Durapatite
  • Mixed Function Oxygenases
  • Vitamin K Epoxide Reductases
  • Epoxide Hydrolases