Comparison of crude and affinity purified cytosolic epoxide hydrolases from hepatic tissue of control and clofibrate-fed mice

Bruce D. Hammock, Glenn D. Prestwich, Dana N. Loury, Peter Y K Cheung, Wai Si Eng, Sang Kyu Park, David E. Moody, Marilyn H. Silva, Roger N. Wixtrom

Research output: Contribution to journalArticle

24 Scopus citations

Abstract

An affinity purification procedure was developed for the cytosolic epoxide hydrolase based upon the selective binding of the enzyme to immobilized methoxycitronellyl thiol. Several elution systems were examined, but the most successful system employed selective elution with a chalcone oxide. This affinity system allowed the purification of the cytosolic epoxide hydrolase activity from livers of both control and clofibrate-fed mice. A variety of biochemical techniques including pH dependence, substrate preference, kinetics, inhibition, amino acid analysis, peptide mapping, Western blotting, analytical isoelectric focusing, and gel permeation chromatography failed to distinguish between the enzymes purified from control and clofibrate-fed animals. The quantitative removal of the cytosolic epoxide hydrolase acting on trans-stilbene oxide from 100,000g supernatants, allowed analysis of remaining activities acting differentially on cis-stilbene oxide and benzo[a]pyrene 4,5-oxide. Such analysis indicated the existence of a novel epoxide hydrolase activity in the cytosol of mouse liver preparations.

Original languageEnglish (US)
Pages (from-to)292-309
Number of pages18
JournalArchives of Biochemistry and Biophysics
Volume244
Issue number1
DOIs
StatePublished - 1986

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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    Hammock, B. D., Prestwich, G. D., Loury, D. N., Cheung, P. Y. K., Eng, W. S., Park, S. K., Moody, D. E., Silva, M. H., & Wixtrom, R. N. (1986). Comparison of crude and affinity purified cytosolic epoxide hydrolases from hepatic tissue of control and clofibrate-fed mice. Archives of Biochemistry and Biophysics, 244(1), 292-309. https://doi.org/10.1016/0003-9861(86)90119-0