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
  • Log out
  • 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
  • Log out
  • 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
Rapid CommunicationShort Communication

Quantitative Transporter Proteomics by Liquid Chromatography with Tandem Mass Spectrometry: Addressing Methodologic Issues of Plasma Membrane Isolation and Expression-Activity Relationship

Vineet Kumar, Bhagwat Prasad, Gabriela Patilea, Anshul Gupta, Laurent Salphati, Raymond Evers, Cornelis E. C. A. Hop and Jashvant D. Unadkat
Drug Metabolism and Disposition February 2015, 43 (2) 284-288; DOI: https://doi.org/10.1124/dmd.114.061614
Vineet Kumar
Department of Pharmaceutics, University of Washington, Seattle, Washington (V.K., B.P., G.P., J.D.U.); Drug Metabolism and Pharmacokinetics, Infection Innovative Medicines Unit, AstraZeneca Pharmaceuticals LLP, Waltham, Massachusetts (A.G.); Pharmacokinetics, Pharmacodynamics and Drug Metabolism, Merck & Co., Kenilworth, New Jersey (R.E.); Drug Metabolism and Pharmacokinetics, Genentech, South San Francisco, Californai (L.S., C.E.C.A.H.)
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Bhagwat Prasad
Department of Pharmaceutics, University of Washington, Seattle, Washington (V.K., B.P., G.P., J.D.U.); Drug Metabolism and Pharmacokinetics, Infection Innovative Medicines Unit, AstraZeneca Pharmaceuticals LLP, Waltham, Massachusetts (A.G.); Pharmacokinetics, Pharmacodynamics and Drug Metabolism, Merck & Co., Kenilworth, New Jersey (R.E.); Drug Metabolism and Pharmacokinetics, Genentech, South San Francisco, Californai (L.S., C.E.C.A.H.)
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Gabriela Patilea
Department of Pharmaceutics, University of Washington, Seattle, Washington (V.K., B.P., G.P., J.D.U.); Drug Metabolism and Pharmacokinetics, Infection Innovative Medicines Unit, AstraZeneca Pharmaceuticals LLP, Waltham, Massachusetts (A.G.); Pharmacokinetics, Pharmacodynamics and Drug Metabolism, Merck & Co., Kenilworth, New Jersey (R.E.); Drug Metabolism and Pharmacokinetics, Genentech, South San Francisco, Californai (L.S., C.E.C.A.H.)
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Anshul Gupta
Department of Pharmaceutics, University of Washington, Seattle, Washington (V.K., B.P., G.P., J.D.U.); Drug Metabolism and Pharmacokinetics, Infection Innovative Medicines Unit, AstraZeneca Pharmaceuticals LLP, Waltham, Massachusetts (A.G.); Pharmacokinetics, Pharmacodynamics and Drug Metabolism, Merck & Co., Kenilworth, New Jersey (R.E.); Drug Metabolism and Pharmacokinetics, Genentech, South San Francisco, Californai (L.S., C.E.C.A.H.)
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Laurent Salphati
Department of Pharmaceutics, University of Washington, Seattle, Washington (V.K., B.P., G.P., J.D.U.); Drug Metabolism and Pharmacokinetics, Infection Innovative Medicines Unit, AstraZeneca Pharmaceuticals LLP, Waltham, Massachusetts (A.G.); Pharmacokinetics, Pharmacodynamics and Drug Metabolism, Merck & Co., Kenilworth, New Jersey (R.E.); Drug Metabolism and Pharmacokinetics, Genentech, South San Francisco, Californai (L.S., C.E.C.A.H.)
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Raymond Evers
Department of Pharmaceutics, University of Washington, Seattle, Washington (V.K., B.P., G.P., J.D.U.); Drug Metabolism and Pharmacokinetics, Infection Innovative Medicines Unit, AstraZeneca Pharmaceuticals LLP, Waltham, Massachusetts (A.G.); Pharmacokinetics, Pharmacodynamics and Drug Metabolism, Merck & Co., Kenilworth, New Jersey (R.E.); Drug Metabolism and Pharmacokinetics, Genentech, South San Francisco, Californai (L.S., C.E.C.A.H.)
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Cornelis E. C. A. Hop
Department of Pharmaceutics, University of Washington, Seattle, Washington (V.K., B.P., G.P., J.D.U.); Drug Metabolism and Pharmacokinetics, Infection Innovative Medicines Unit, AstraZeneca Pharmaceuticals LLP, Waltham, Massachusetts (A.G.); Pharmacokinetics, Pharmacodynamics and Drug Metabolism, Merck & Co., Kenilworth, New Jersey (R.E.); Drug Metabolism and Pharmacokinetics, Genentech, South San Francisco, Californai (L.S., C.E.C.A.H.)
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Jashvant D. Unadkat
Department of Pharmaceutics, University of Washington, Seattle, Washington (V.K., B.P., G.P., J.D.U.); Drug Metabolism and Pharmacokinetics, Infection Innovative Medicines Unit, AstraZeneca Pharmaceuticals LLP, Waltham, Massachusetts (A.G.); Pharmacokinetics, Pharmacodynamics and Drug Metabolism, Merck & Co., Kenilworth, New Jersey (R.E.); Drug Metabolism and Pharmacokinetics, Genentech, South San Francisco, Californai (L.S., C.E.C.A.H.)
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • Article
  • Figures & Data
  • Info & Metrics
  • eLetters
  • PDF + SI
  • PDF
Loading

Abstract

To predict transporter-mediated drug disposition using physiologically based pharmacokinetic models, one approach is to measure transport activity and relate it to protein expression levels in cell lines (overexpressing the transporter) and then scale these to via in vitro to in vivo extrapolation (IVIVE). This approach makes two major assumptions. First, that the expression of the transporter is predominantly in the plasma membrane. Second, that there is a linear correlation between expression level and activity of the transporter protein. The present study was conducted to test these two assumptions. We evaluated two commercially available kits that claimed to separate plasma membrane from other cell membranes. The Qiagen Qproteome kit yielded very little protein in the fraction purported to be the plasma membrane. The Abcam Phase Separation kit enriched the plasma membrane but did not separate it from other intracellular membranes. For the Abcam method, the expression level of organic anion-transporting polypeptides (OATP) 1B1/2B1 and breast cancer resistance protein (BCRP) proteins in all subcellular fractions isolated from cells or human liver tissue tracked that of Na+-K+ ATPase. Assuming that Na+-K+ ATPase is predominantly located in the plasma membrane, these data suggest that the transporters measured are also primarily located in the plasma membrane. Using short hairpin RNA, we created clones of cell lines with varying degrees of OATP1B1 or BCRP expression level. In these clones, transport activity of OATP1B1 or BCRP was highly correlated with protein expression level (r2 > 0.9). These data support the use of transporter expression level data and activity data from transporter overexpressing cell lines for IVIVE of transporter-mediated disposition of drugs.

Footnotes

    • Received October 17, 2014.
    • Accepted December 8, 2014.
  • A.G., L.S., R.E., and C.E.C.A.H. contributed equally to this work.

  • This study was supported by the University of Washington Research Affiliate Program on Transporters (UWRAPT) sponsored by AstraZeneca, Genentech, and Merck & Co., Inc. (http://sop.washington.edu/department-of-pharmaceutics/research-affiliate-program-on-transporters-uwrapt/).

  • dx.doi.org/10.1124/dmd.114.061614.

  • ↵Embedded ImageThis article has supplemental material available at dmd.aspetjournals.org.

  • Copyright © 2014 by The American Society for Pharmacology and Experimental Therapeutics
View Full Text

 

DMD articles become freely available 12 months after publication, and remain freely available for 5 years. 

Non-open access articles that fall outside this five year window are available only to institutional subscribers and current ASPET members, or through the article purchase feature at the bottom of the page. 

 

  • Click here for information on institutional subscriptions.
  • Click here for information on individual ASPET membership.

 

Log in using your username and password

Forgot your user name or password?

Purchase access

You may purchase access to this article. This will require you to create an account if you don't already have one.
PreviousNext
Back to top

In this issue

Drug Metabolism and Disposition: 43 (2)
Drug Metabolism and Disposition
Vol. 43, Issue 2
1 Feb 2015
  • 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.
Quantitative Transporter Proteomics by Liquid Chromatography with Tandem Mass Spectrometry: Addressing Methodologic Issues of Plasma Membrane Isolation and Expression-Activity Relationship
(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
Rapid CommunicationShort Communication

Transporter Expression-Activity Correlation

Vineet Kumar, Bhagwat Prasad, Gabriela Patilea, Anshul Gupta, Laurent Salphati, Raymond Evers, Cornelis E. C. A. Hop and Jashvant D. Unadkat
Drug Metabolism and Disposition February 1, 2015, 43 (2) 284-288; DOI: https://doi.org/10.1124/dmd.114.061614

Citation Manager Formats

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

Share
Rapid CommunicationShort Communication

Transporter Expression-Activity Correlation

Vineet Kumar, Bhagwat Prasad, Gabriela Patilea, Anshul Gupta, Laurent Salphati, Raymond Evers, Cornelis E. C. A. Hop and Jashvant D. Unadkat
Drug Metabolism and Disposition February 1, 2015, 43 (2) 284-288; DOI: https://doi.org/10.1124/dmd.114.061614
Reddit logo Twitter logo Facebook logo Mendeley logo
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Jump to section

  • Article
    • Abstract
    • Introduction
    • Materials and Methods
    • Results
    • Discussion
    • Acknowledgments
    • Authorship Contributions
    • Footnotes
    • Abbreviations
    • References
  • Figures & Data
  • Info & Metrics
  • eLetters
  • PDF + SI
  • PDF

Related Articles

Cited By...

More in this TOC Section

  • Preincubation Effects on Inhibition of OCT1 by CsA
  • Carbamazepine Metabolite and Hypersensitivity Reactions
  • SULT4A1 Preserves Mitochondrial Function
Show more Short Communications

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