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

Structure-function analysis of human CYP3A4 using a specific proinhibitory antipeptide antibody.

T Schulz-Utermoehl1, R J Mountfield, R P Bywater

  • 1Department of Drug Metabolism, Novo Nordisk A/S, Maaloev, Denmark. tisu@novo.dk

Drug Metabolism and Disposition: the Biological Fate of Chemicals
|June 20, 2000
PubMed
Summary

An antibody targeting human CYP3A4 (cytochrome P450 3A4) potently inhibited its activity. This research reveals the targeted loop region is crucial for catalysis, not substrate binding, impacting enzyme efficiency.

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

Improved Pregnancy Outcomes in Beef Heifers Through Delayed Insemination of Sexed Semen.

Reproduction in domestic animals = Zuchthygiene·2025
Same author

NSGO-OV-UMB1/ENGOT-OV30: A phase II study of durvalumab in combination with the anti-CD73 monoclonal antibody Oleclumab in patients with relapsed ovarian cancer.

Gynecologic oncology·2024
Same author

Circulating immune-related proteins associated with immune checkpoint inhibitor efficacy in patients with pancreatic ductal adenocarcinoma.

ESMO open·2024
Same author

Association between the length of in vitro embryo culture, mode of ART, and the initial endogenous hCG rise in ongoing singleton pregnancies.

Human reproduction (Oxford, England)·2024
Same author

Metastatic atypical renal tumour with metanephric characteristics treated with Sunitinib.

Urology case reports·2021
Same author

Associations between community programmes and policies and children's physical activity: the Healthy Communities Study.

Pediatric obesity·2018

Area of Science:

  • Biochemistry
  • Enzymology
  • Drug Metabolism

Background:

  • Human CYP3A4 is a critical enzyme in drug metabolism.
  • Understanding the catalytic mechanism of CYP3A4 is essential for drug development and safety.
  • Specific regions of CYP3A4 play vital roles in its enzymatic activity.

Purpose of the Study:

  • To investigate the function of residues 253-269 of human CYP3A4 in its catalytic activity.
  • To determine the role of this specific region in substrate binding, electron transfer, and oxygen activation during catalysis.

Main Methods:

  • Production of an anti-peptide antibody targeting human CYP3A4 residues 253-269.
  • Assay of CYP3A4 activity using testosterone as a substrate in human hepatic microsomal fractions.
  • Spectroscopic analysis (Type I spectrum) and kinetic studies (K(m), V(max)) were performed.

Related Experiment Videos

  • Investigation of P450-CO complex formation and NADPH oxidation.

    • Main Results:

    • The antibody specifically and potently inhibited CYP3A4 activity (>90%).
    • Antibody binding did not affect testosterone binding (Type I spectrum) or K(m) but significantly decreased V(max) (>90%).
    • NADPH oxidation was reduced by 60% in the presence of the antibody.

    Conclusions:

    • The targeted loop region (residues 253-269) is not involved in substrate binding, reductase interaction, electron transfer, or oxygen binding.
    • Antibody binding likely inhibits CYP3A4 activity by destabilizing the hydroperoxo complex, interfering with proton transfer, or hindering substrate-induced conformational changes.