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

Human CYP2B6: expression, inducibility and catalytic activities.

L Gervot1, B Rochat, J C Gautier

  • 1INSERM U490, Université René Descartes, Paris, France.

Pharmacogenetics
|September 2, 1999
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

Tobacco cessation and the role of ESMO and medical oncologists: addressing the specific needs of cancer patients in times of the COVID-19 pandemic.

ESMO open·2023
Same author

Corrigendum to "Validation and clinical application of a multiplex high performance liquid chromatography - Tandem mass spectrometry assay for the monitoring of plasma concentrations of 12 antibiotics in patients with severe bacterial infections" [J. Chromatogr. B 1157 (2020) 122160].

Journal of chromatography. B, Analytical technologies in the biomedical and life sciences·2022
Same author

Validation and clinical application of a multiplex high performance liquid chromatography - tandem mass spectrometry assay for the monitoring of plasma concentrations of 12 antibiotics in patients with severe bacterial infections.

Journal of chromatography. B, Analytical technologies in the biomedical and life sciences·2020
Same author

Effects of oral supplementation of iron on hepcidin blood concentrations among non-anaemic female blood donors: a randomized controlled trial.

Vox sanguinis·2015
Same author

Quasi-electric fields and band offsets: teaching electrons new tricks (Nobel lecture).

Chemphyschem : a European journal of chemical physics and physical chemistry·2013
Same author

Importance of influx and efflux systems and xenobiotic metabolizing enzymes in intratumoral disposition of anticancer agents.

Current cancer drug targets·2009
Same journal

No evidence of a role for PPARgamma Pro12Ala polymorphism in endometrial cancer susceptibility.

Pharmacogenetics·2004
Same journal

Pharmacogenetics and herb-drug interactions: experience with Ginkgo biloba and omeprazole.

Pharmacogenetics·2004
Same journal

Functional comparison of the endothelial nitric oxide synthase Glu298Asp polymorphic variants in human endothelial cells.

Pharmacogenetics·2004
Same journal

Response to micronized fenofibrate treatment is associated with the peroxisome-proliferator-activated receptors alpha G/C intron7 polymorphism in subjects with type 2 diabetes.

Pharmacogenetics·2004
Same journal

Upstream and coding region CYP2C9 polymorphisms: correlation with warfarin dose and metabolism.

Pharmacogenetics·2004
Same journal

The human serotonin receptor 2B: coding region polymorphisms and association with vulnerability to illegal drug abuse.

Pharmacogenetics·2004
See all related articles

Researchers expressed active human cytochrome P450 2B6 (CYP2B6) in yeast and bacteria. This allowed for detailed studies on CYP2B6 metabolism, expression, and inducibility in various human tissues and cells.

Area of Science:

  • Biochemistry
  • Pharmacology
  • Molecular Biology

Background:

  • Cytochrome P450 2B6 (CYP2B6) is a key enzyme in drug metabolism.
  • Understanding CYP2B6 expression and activity is crucial for personalized medicine and drug development.

Purpose of the Study:

  • To express active human CYP2B6 in microbial systems for functional and immunological studies.
  • To characterize the substrate specificity and metabolic activity of recombinant CYP2B6.
  • To develop a specific antibody for investigating CYP2B6 expression and inducibility in human tissues and cells.

Main Methods:

  • Cloning and expression of human CYP2B6 cDNA in Saccharomyces cerevisiae and Escherichia coli.
  • Enzymatic assays using various substrates (ethoxyresorufin, pentoxyresorufin, benzyloxyresorufin, ethoxycoumarin, testosterone, cyclophosphamide).

Related Experiment Videos

  • Production of a specific anti-human CYP2B6 antibody in E. coli.
  • Immunoblot analysis and mRNA detection in human liver samples, extrahepatic tissues, and cell lines.
  • Assessment of CYP2B6 induction by phenobarbital and cyclophosphamide.

    • Main Results:

    • High-level expression of active CYP2B6 in yeast, enabling kinetic analysis.
    • CYP2B6 preferentially metabolized benzyloxyresorufin and pentoxyresorufin, and showed strong 4-hydroxycyclophosphamide activity.
    • A highly specific anti-CYP2B6 antibody was generated, with no cross-reactivity against other major CYP isoforms.
    • CYP2B6 protein was detected in 43/48 human liver samples and in brain, intestine, kidney, and lung.
    • CYP2B6 mRNA was found in liver, kidney, lung, trachea, and intestine.
    • CYP2B6 expression was induced at both protein and mRNA levels by phenobarbital and cyclophosphamide in primary human hepatocytes.

    Conclusions:

    • Recombinant CYP2B6 expressed in yeast is enzymatically active and useful for substrate specificity studies.
    • The specific antibody is a valuable tool for studying CYP2B6 expression and distribution in humans.
    • CYP2B6 is expressed in various human tissues and is inducible by phenobarbital and cyclophosphamide, highlighting its clinical relevance.