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
Molecular Pharmacology
  • 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
Molecular Pharmacology

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 molpharm on Facebook
  • Follow molpharm on Twitter
  • Follow molpharm on LinkedIn
Research ArticleArticle

Reconstitution of the Human 5-HT1D Receptor-G-Protein Coupling: Evidence for Constitutive Activity and Multiple Receptor Conformations

Reginald Brys, Katty Josson, Maria Paola Castelli, Mirek Jurzak, Peter Lijnen, Walter Gommeren and Josée E. Leysen
Molecular Pharmacology June 2000, 57 (6) 1132-1141;
Reginald Brys
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Katty Josson
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Maria Paola Castelli
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Mirek Jurzak
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Peter Lijnen
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Walter Gommeren
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Josée E. Leysen
  • 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
Loading

Abstract

The 5-hydroxytryptamine (5-HT) 1D/1B receptors have gained particular interest as potential targets for treatment of migraine and depression. G-protein coupling and other intrinsic properties of the human 5-HT1D receptor were studied using a baculovirus-based expression system in Sf9 cells. Coexpression of the human 5-HT1D receptor with Gαi1, αi2, αi3, or Gαo-proteins and Gβ1γ2-subunits reconstituted a Gpp(NH)p-sensitive, high affinity binding of [3H]5-HT to this receptor, whereas the Gαqβ1γ2 heterotrimer was ineffective in this respect. Competition of [3H]5-HT binding by various compounds confirmed that coexpression of the human 5-HT1D receptor with Gαi/oβ1γ2 reconstitutes the receptor in a high affinity agonist binding state, having the same pharmacological profile as the receptor expressed in mammalian cells. Binding of the antagonist ocaperidone to the human 5-HT1Dreceptor in coupled or noncoupled state was analyzed. This compound competed with [3H]5-HT binding more potently on the human 5-HT1D receptor in the noncoupled state, showing its inverse agonistic character. Ocaperidone acted as a competitive inhibitor of [3H]5-HT binding when tested with the coupled receptor form but not so when tested with the noncoupled receptor preparation. Finally, [35S]GTPγS binding experiments using the inverse agonist ocaperidone revealed a high level of constitutive activity of the human 5-HT1D receptor. Taken together, the reconstitution of the human 5-HT1Dreceptor-G-protein coupling using baculovirus-infected Sf9 cells made possible the assessment of coupling specificity and the detection of different binding states of the receptor induced by G-protein coupling or ligand binding.

Footnotes

  • Send reprint requests to: Josée Leysen, Janssen Research Foundation, Turnhoutseweg 30, B-2340 Beerse, Belgium. E-mail:jleysen2{at}janbe.jnj.com

  • ↵1 These authors contributed equally to the work.

  • This work was supported in part by the IWT Grant 940232.

  • Abbreviations:
    5-HT
    5-hydroxytryptamine
    R
    receptor
    Gαi/o
    combination of Gαi1, Gαi2, Gαi3, and Gαo
    GPCR
    G-protein-coupled receptor
    Gpp(NH)p
    guanosine 5′-(β,γ-imido)triphosphate
    GTPγS
    guanosine 5′-O-(3-thiotriphosphate)
    Sf9 cells
    Spodoptera frugiperda cells
    pIC50
    −log of the IC50 value
    pEC50
    −log of the EC50, value
    • Received July 19, 1999.
    • Accepted February 20, 2000.
  • The American Society for Pharmacology and Experimental Therapeutics
View Full Text

MolPharm 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

Molecular Pharmacology: 57 (6)
Molecular Pharmacology
Vol. 57, Issue 6
1 Jun 2000
  • Table of Contents
  • About the Cover
  • Index by author
Download PDF
Article Alerts
Sign In to Email Alerts with your Email Address
Email Article

Thank you for sharing this Molecular Pharmacology 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.
Reconstitution of the Human 5-HT1D Receptor-G-Protein Coupling: Evidence for Constitutive Activity and Multiple Receptor Conformations
(Your Name) has forwarded a page to you from Molecular Pharmacology
(Your Name) thought you would be interested in this article in Molecular Pharmacology.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Citation Tools
Research ArticleArticle

Reconstitution of the Human 5-HT1D Receptor-G-Protein Coupling: Evidence for Constitutive Activity and Multiple Receptor Conformations

Reginald Brys, Katty Josson, Maria Paola Castelli, Mirek Jurzak, Peter Lijnen, Walter Gommeren and Josée E. Leysen
Molecular Pharmacology June 1, 2000, 57 (6) 1132-1141;

Citation Manager Formats

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

Share
Research ArticleArticle

Reconstitution of the Human 5-HT1D Receptor-G-Protein Coupling: Evidence for Constitutive Activity and Multiple Receptor Conformations

Reginald Brys, Katty Josson, Maria Paola Castelli, Mirek Jurzak, Peter Lijnen, Walter Gommeren and Josée E. Leysen
Molecular Pharmacology June 1, 2000, 57 (6) 1132-1141;
Reddit logo Twitter logo Facebook logo Mendeley logo
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Jump to section

  • Article
    • Abstract
    • Experimental Procedures
    • Results
    • Discussion
    • Acknowledgments
    • Footnotes
    • References
  • Figures & Data
  • Info & Metrics
  • eLetters
  • PDF

Related Articles

Cited By...

More in this TOC Section

  • Antimicrobial and Antileukemic Transportan 10 Conjugates
  • Pharmacological characterization of zebrafish H1 receptor
  • Bhave and Forman
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 Molecular Pharmacology
  • Editorial Board
  • Instructions to Authors
  • Submit a Manuscript
  • Customized Alerts
  • RSS Feeds
  • Subscriptions
  • Permissions
  • Terms & Conditions of Use

ASPET's Other Journals

  • Drug Metabolism and Disposition
  • Journal of Pharmacology and Experimental Therapeutics
  • Pharmacological Reviews
  • Pharmacology Research & Perspectives
ISSN 1521-0111 (Online)

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