Skip to main content
Advertisement

Main menu

  • Home
  • Articles
    • Current Issue
    • Fast Forward
    • Latest Articles
    • Special Sections
    • Archive
  • Information
    • Instructions to Authors
    • Submit a Manuscript
    • FAQs
    • For Subscribers
    • Terms & Conditions of Use
    • Permissions
  • Editorial Board
  • Alerts
    • Alerts
    • RSS Feeds
  • Virtual Issues
  • Feedback
  • Submit
  • Other Publications
    • Drug Metabolism and Disposition
    • Journal of Pharmacology and Experimental Therapeutics
    • Molecular Pharmacology
    • Pharmacological Reviews
    • Pharmacology Research & Perspectives
    • ASPET

User menu

  • My alerts
  • Log in
  • My Cart

Search

  • Advanced search
Drug Metabolism & Disposition
  • Other Publications
    • Drug Metabolism and Disposition
    • Journal of Pharmacology and Experimental Therapeutics
    • Molecular Pharmacology
    • Pharmacological Reviews
    • Pharmacology Research & Perspectives
    • ASPET
  • My alerts
  • Log in
  • My Cart
Drug Metabolism & Disposition

Advanced Search

  • Home
  • Articles
    • Current Issue
    • Fast Forward
    • Latest Articles
    • Special Sections
    • Archive
  • Information
    • Instructions to Authors
    • Submit a Manuscript
    • FAQs
    • For Subscribers
    • Terms & Conditions of Use
    • Permissions
  • Editorial Board
  • Alerts
    • Alerts
    • RSS Feeds
  • Virtual Issues
  • Feedback
  • Submit
  • Visit dmd on Facebook
  • Follow dmd on Twitter
  • Follow ASPET on LinkedIn
OtherArticle

Quantification of accurate composition and total abundance of homologous proteins by conserved-plus-surrogate peptide (CPSP) approach: Quantification of UDP glucuronosyltransferases in human tissues

Deepak Ahire, Mitesh Patel, Sujal V Deshmukh and Bhagwat Prasad
Drug Metabolism and Disposition November 29, 2022, DMD-AR-2022-001155; DOI: https://doi.org/10.1124/dmd.122.001155
Deepak Ahire
1Department of Pharmaceutical Sciences, Washington State University, United States
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Deepak Ahire
Mitesh Patel
2Novartis Institutes for BioMedical Research, United States
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Sujal V Deshmukh
2Novartis Institutes for BioMedical Research, United States
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Bhagwat Prasad
1Department of Pharmaceutical Sciences, Washington State University, United States
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Bhagwat Prasad
  • For correspondence: bhagwat.prasad@wsu.edu
  • Article
  • Figures & Data
  • Info & Metrics
  • eLetters
  • PDF + SI
  • PDF
Loading

Abstract

Characterization of accurate compositions and total abundance of homologous drug-metabolizing enzymes, such as UDP glucuronosyltransferases (UGTs), is important for predicting the fractional contribution of individual isoforms involved in the metabolism of a drug for applications in physiologically based pharmacokinetic (PBPK) modeling. Conventional targeted proteomics utilizes surrogate peptides, which often results in high technical and inter-laboratory variability due to peptide-specific digestion efficiency leading to data inconsistencies. To address this problem, we developed a novel universal conserved-plus-surrogate peptide (CPSP) approach for determining the accurate compositions and total or cumulative abundance of homologous UGTs in commercially available pooled human liver microsomes (HLM), human intestinal microsomes (HIM), human kidney microsomes (HKM), and human liver S9 (HLS9) fractions. The relative percent composition of UGT1A and UGT2B isoforms in human liver was 35:5:36:11:13 for UGT1A1:1A3:1A4:1A6:1A9, and 20:32:22:21:5 for UGT2B4:2B7:2B10:2B15:2B17. The human kidney and intestine also show unique compositions of UGT1As and UGT2Bs. The reproducibility of the approach was validated by assessing correlations of UGT compositions between HLM and HLS9 (R2>0.91). The analysis of conserved peptides also provided the absolute abundance for individual UGT isoforms included in this investigation as well as the total abundance (pmol/mg protein) of UGT1As and UGT2Bs across tissues, i.e., 268 and 342 (HLM), 21 and 92 (HIM), 138 and 99 (HKM), respectively. In summary, the CPSP approach could be utilized for applications in the in-vitro to in-vivo extrapolation (IVIVE) of drug metabolism and PBPK modeling.

Significance Statement We quantified the absolute compositions and total abundance of UGTs in human liver, intestine, and kidney microsomes using a novel CPSP approach. Unlike the conventional surrogate peptide-based targeted proteomics, the CPSP approach addresses the surrogate peptide-specific variability in the determination of the absolute composition of UGTs. The data presented in this manuscript are applicable for the estimation of the fraction metabolized (fm) by individual UGTs towards better IVIVE of UGT-mediated drug metabolism in the human liver, intestine, and kidney.

  • drug metabolism
  • pharmacokinetic modeling
  • phase II drug metabolism
  • UDP glucuronosyltransferase (UGT)
  • Copyright © 2020 American Society for Pharmacology and Experimental Therapeutics
PreviousNext
Back to top

In this issue

Drug Metabolism and Disposition: 51 (2)
Drug Metabolism and Disposition
Vol. 51, Issue 2
1 Feb 2023
  • Table of Contents
  • Table of Contents (PDF)
  • About the Cover
  • Index by author
  • Editorial Board (PDF)
  • Front Matter (PDF)
Download PDF
Article Alerts
Sign In to Email Alerts with your Email Address
Email Article

Thank you for sharing this Drug Metabolism & Disposition article.

NOTE: We request your email address only to inform the recipient that it was you who recommended this article, and that it is not junk mail. We do not retain these email addresses.

Enter multiple addresses on separate lines or separate them with commas.
Quantification of accurate composition and total abundance of homologous proteins by conserved-plus-surrogate peptide (CPSP) approach: Quantification of UDP glucuronosyltransferases in human tissues
(Your Name) has forwarded a page to you from Drug Metabolism & Disposition
(Your Name) thought you would be interested in this article in Drug Metabolism & Disposition.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Citation Tools
OtherArticle

A novel CPSP approach for quantification of UGT enzymes

Deepak Ahire, Mitesh Patel, Sujal V Deshmukh and Bhagwat Prasad
Drug Metabolism and Disposition November 29, 2022, DMD-AR-2022-001155; DOI: https://doi.org/10.1124/dmd.122.001155

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero

Share
OtherArticle

A novel CPSP approach for quantification of UGT enzymes

Deepak Ahire, Mitesh Patel, Sujal V Deshmukh and Bhagwat Prasad
Drug Metabolism and Disposition November 29, 2022, DMD-AR-2022-001155; DOI: https://doi.org/10.1124/dmd.122.001155
del.icio.us logo Digg logo Reddit logo Twitter logo Facebook logo Google logo Mendeley logo
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Jump to section

  • Article
  • Figures & Data
  • Info & Metrics
  • eLetters
  • PDF + SI
  • PDF

Related Articles

Cited By...

More in this TOC Section

  • Series-Compartment Models of Hepatic Elimination
  • Warfarin PBPK Model with TMDD Mechanism
  • Identification of payload-containing catabolites of ADCs
Show more Article

Similar Articles

Advertisement
  • Home
  • Alerts
Facebook   Twitter   LinkedIn   RSS

Navigate

  • Current Issue
  • Fast Forward by date
  • Fast Forward by section
  • Latest Articles
  • Archive
  • Search for Articles
  • Feedback
  • ASPET

More Information

  • About DMD
  • Editorial Board
  • Instructions to Authors
  • Submit a Manuscript
  • Customized Alerts
  • RSS Feeds
  • Subscriptions
  • Permissions
  • Terms & Conditions of Use

ASPET's Other Journals

  • Journal of Pharmacology and Experimental Therapeutics
  • Molecular Pharmacology
  • Pharmacological Reviews
  • Pharmacology Research & Perspectives
ISSN 1521-009X (Online)

Copyright © 2023 by the American Society for Pharmacology and Experimental Therapeutics