Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Liver microsomal expoxide hydrase. Solubilization, purification, and characterization.

A Y Lu, D Ryan, D M Jerina

    The Journal of Biological Chemistry
    |October 25, 1975
    PubMed
    Summary
    This summary is machine-generated.

    Related Concept Videos

    You might also read

    Related Articles

    Articles linked to this work by shared authors, journal, and citation graph.

    Sort by
    Same author

    Brainstem melanomas presenting as a cavernous malformation.

    Neuro-Chirurgie·2014
    Same author

    Metabolism of benzo(e)pyrene by rat liver microsomal enzymes.

    Carcinogenesis·2012
    Same author

    Covalent binding of chemical residues: health impact.

    Advances in experimental medicine and biology·2002
    Same author

    Dose-dependent differences in the profile of mutations induced by carcinogenic (R,S,S,R) bay- and fjord-region diol epoxides of polycyclic aromatic hydrocarbons.

    Advances in experimental medicine and biology·2002
    Same author

    Patterns of resistance to exonuclease digestion of oligonucleotides containing polycyclic aromatic hydrocarbon diol epoxide adducts at N6 of deoxyadenosine.

    Chemical research in toxicology·2001
    Same author

    Cross-linking of the fingers subdomain of human immunodeficiency virus type 1 reverse transcriptase to template-primer.

    Journal of virology·2001

    Researchers purified epoxide hydrase (EH) from rat liver microsomes. The purified enzyme, a key component in xenobiotic metabolism, showed a molecular weight of 53-54 kDa.

    Area of Science:

    • Biochemistry
    • Enzymology
    • Drug Metabolism

    Background:

    • Epoxide hydrase (EH) is crucial for metabolizing epoxides, reactive intermediates in xenobiotic metabolism.
    • Phenobarbital treatment induces EH activity in rat liver microsomes.
    • Understanding EH purification is vital for studying its role in detoxification.

    Purpose of the Study:

    • To solubilize and purify epoxide hydrase from liver microsomes of phenobarbital-treated rats.
    • To characterize the molecular properties of the purified enzyme.

    Main Methods:

    • Solubilization using cholate.
    • Purification via ammonium sulfate fractionation and column chromatography (DEAE-cellulose, hydroxylapatite) with Emulgen 911.
    • Molecular weight determination using SDS-polyacrylamide gel electrophoresis.

    Related Experiment Videos

    Main Results:

    • Apparent homogeneity of the purified epoxide hydrase.
    • A single major band of 53,000-54,000 molecular weight under SDS-PAGE.
    • Evidence of high molecular weight aggregates in the absence of SDS.
    • Preparation was largely free of heme and flavin, but contained residual lipids and Emulgen 911.

    Conclusions:

    • A method for purifying epoxide hydrase to apparent homogeneity was established.
    • The purified enzyme exists as aggregates in solution but monomers of 53-54 kDa under denaturing conditions.
    • The purified preparation provides a basis for further biochemical and mechanistic studies of epoxide hydrase.