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Assaying Surface Expression of Chemosensory Receptors in Heterologous Cells
04:55

Assaying Surface Expression of Chemosensory Receptors in Heterologous Cells

Published on: February 23, 2011

Olfactory receptor surface expression is driven by association with the beta2-adrenergic receptor

Chris Hague1, Michelle A Uberti, Zhongjian Chen

  • 1Department of Pharmacology, Emory University School of Medicine, Atlanta, GA 30322, USA. chague@emory.edu

Proceedings of the National Academy of Sciences of the United States of America
|September 7, 2004
PubMed

Insights

Co-expressing olfactory receptors (ORs) with beta(2)-adrenergic receptors (ARs) significantly enhances OR surface expression. This physical interaction allows for functional studies of wild-type ORs, advancing olfactory receptor research.

Area of Science:

  • Biochemistry
  • Neuroscience
  • Molecular Biology

Background:

  • Olfactory receptors (ORs) are a major class of G protein-coupled receptors (GPCRs).
  • Wild-type ORs exhibit poor cell surface expression in heterologous systems, hindering functional studies.
  • Receptor-receptor interactions are known to influence GPCR trafficking.

Purpose of the Study:

  • To investigate if interactions with other GPCRs can enhance olfactory receptor surface expression.
  • To determine the role of beta-adrenergic receptor (AR) subtypes in OR trafficking.
  • To demonstrate functional expression of a wild-type OR in a heterologous system.

Main Methods:

  • Coexpression of mouse 71 (M71) OR with various beta-AR subtypes in HEK293 cells.
  • Coimmunoprecipitation and cointernalization assays to assess receptor interaction.
  • Ligand-induced cAMP assays to measure functional activity.
  • In situ hybridization to examine receptor colocalization in olfactory epithelium.

Main Results:

  • Coexpression with beta(2)-ARs, but not other AR subtypes, dramatically increased M71 OR surface expression.
  • A persistent physical association between M71 OR and beta(2)-AR was confirmed.
  • Coexpression enabled functional cAMP responses to the M71 ligand, acetophenone.
  • Extensive colocalization of M71 OR and beta(2)-AR was observed in mouse olfactory epithelium.

Conclusions:

  • Successful heterologous surface expression of a functional wild-type OR is achieved.
  • Persistent physical association with beta(2)-ARs controls OR surface expression.
  • This interaction provides a novel strategy for studying olfactory receptor function.

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