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

Differential Signaling by Splice Variants of the Human Free Fatty Acid Receptor GPR120

Sarah-Jane Watson, Alastair J. H. Brown and Nicholas D. Holliday
Molecular Pharmacology May 2012, 81 (5) 631-642; DOI: https://doi.org/10.1124/mol.111.077388
Sarah-Jane Watson
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Alastair J. H. Brown
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Nicholas D. Holliday
  • 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

GPR120 is a long-chain fatty acid receptor that stimulates incretin hormone release from colonic endocrine cells and is implicated in macrophage and adipocyte function. The functional consequences of long (L) and short (S) human GPR120 splice variants, which differ by insertion of 16 amino acids in the third intracellular loop, are currently unknown. Here we compare signaling and intracellular trafficking of GPR120S and GPR120L receptors, using calcium mobilization and dynamic mass redistribution (DMR) assays, together with quantitative imaging measurements of β-arrestin2 association and receptor internalization. FLAG- or SNAP-tagged GPR120S receptors elicited both intracellular calcium mobilization and DMR responses in human embryonic kidney 293 cells, when stimulated with oleic acid, myristic acid, or the agonist 4-[[(3-phenoxyphenyl)methyl]amino]benzenepropanoic acid (GW9508). Responses were insensitive to pertussis toxin, but increases in intracellular calcium were attenuated by 2-aminoethoxydiphenyl borate, an inhibitor of store inositol trisphosphate receptors. Despite equivalent cell surface expression of SNAP-tagged GPR120L receptors, no specific calcium or DMR responses were observed in cells transfected with this isoform. However, agonist-stimulated GPR120S and GPR120L receptors both recruited β-arrestin2 and underwent robust internalization, with similar agonist potencies in each case. After oleic acid-induced internalization, neither GPR120 isoform recycled rapidly to the cell surface. In both cases, confocal microscopy indicated receptor targeting to lysosomal compartments. Thus, the third intracellular loop insertion in GPR120L prevents G protein-dependent intracellular calcium and DMR responses, but this receptor isoform remains functionally coupled to the β-arrestin pathway, providing one of the first examples of a native β-arrestin-biased receptor.

Footnotes

  • ↵Embedded Image The online version of this article (available at http://molpharm.aspetjournals.org) contains supplemental material.

  • This work was supported by AstraZeneca (Alderley Park, UK); the UK Medical Research Council [Grant G0700049] (to N.D.H.); and the UK Engineering and Physical Sciences Research Council (postgraduate studentship to S.J.W.).

  • Article, publication date, and citation information can be found at http://molpharm.aspetjournals.org.

    http://dx.doi.org/10.1124/mol.111.077388.

  • ABBREVIATIONS:

    FFA
    free fatty acid
    PPAR
    peroxisome proliferator-activated receptor
    GPCR
    G protein-coupled receptor
    ICL
    intracellular loop
    HEK
    human embryonic kidney
    GW9508
    (4-[[(3-phenoxyphenyl)methyl]amino]benzenepropanoic acid)
    DMR
    dynamic mass redistribution
    DMSO
    dimethyl sulfoxide
    PTX
    pertussis toxin
    TR
    tetracycline repressor
    GFP
    green fluorescent protein
    BiFC
    bimolecular fluorescence complementation
    DMEM
    Dulbecco's modified Eagle's medium
    FBS
    fetal bovine serum
    HBSS
    HEPES buffered saline solution
    BSA
    bovine serum albumin
    BG
    benzyl guanine
    BG-AF488
    benzyl guanine-Alexa Fluor-488
    H33342
    Hoechst 33342
    BSA
    bovine serum albumin
    OA
    oleic acid
    Myr
    myristic acid
    2-APB
    2-aminoethoxydiphenyl borate
    TMD
    transmembrane domain
    TZD
    thiazolidinedione.

  • Received December 21, 2011.
  • Accepted January 26, 2012.
  • Copyright © 2012 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: 81 (5)
Molecular Pharmacology
Vol. 81, Issue 5
1 May 2012
  • 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 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.
Differential Signaling by Splice Variants of the Human Free Fatty Acid Receptor GPR120
(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

Signaling Specificity of Human GPR120 Isoforms

Sarah-Jane Watson, Alastair J. H. Brown and Nicholas D. Holliday
Molecular Pharmacology May 1, 2012, 81 (5) 631-642; DOI: https://doi.org/10.1124/mol.111.077388

Citation Manager Formats

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

Share
Research ArticleArticle

Signaling Specificity of Human GPR120 Isoforms

Sarah-Jane Watson, Alastair J. H. Brown and Nicholas D. Holliday
Molecular Pharmacology May 1, 2012, 81 (5) 631-642; DOI: https://doi.org/10.1124/mol.111.077388
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
    • Abstract
    • Introduction
    • Materials and Methods
    • Results
    • Discussion
    • Authorship Contributions
    • Acknowledgments
    • Footnotes
    • References
  • Figures & Data
  • Info & Metrics
  • eLetters
  • PDF + SI
  • PDF

Related Articles

Cited By...

More in this TOC Section

  • Therapeutic Effects of FGF23 Antagonists in Hyp Mice
  • TRPV3 and TRPV4 Channels Coassemble into Heterotetramers
  • Secretin Amino-Terminal Structure-Activity Relationships and Complementary Mutagenesis at the Site of Docking to the Secretin Receptor
Show more Articles

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 © 2022 by the American Society for Pharmacology and Experimental Therapeutics