A Robust, Rapid, and Simple Method of Producing Olfactory Receptors Using Commercial E. coli Cell-Free Systems

  • Karolina Corin
  • Brian Cook
  • Shuguang Zhang
Part of the Methods in Molecular Biology book series (MIMB, volume 1003)


The first bottleneck in olfactory receptor (OR) studies is producing sufficient quantities of soluble, ­functional, and stable receptors. Commercial cell-free in vitro translation systems can be used to produce milligrams of soluble and functional receptors within several hours directly from plasmid DNA. The receptors can be purified using immunoaffinity chromatography and gel filtration, and can be analyzed using gel electrophoresis and with other standard techniques.

Key words

In vitro translation Olfactory receptors Odorant ligand-binding Microscale thermophoresis 


  1. 1.
    Grisshammer R, Duckworth R, Henderson R (1993) Expression of a rat neurotensin receptor in Escherichia coli. Biochem J 295:571–576PubMedGoogle Scholar
  2. 2.
    Hampe W et al (2000) Engineering of a proteolytically stable human β2-adrenergic receptor/maltose-binding protein fusion and production of the chimeric protein in Escherichia coli and baculovirus infected insect cells. J Biotechnol 77:219–234PubMedCrossRefGoogle Scholar
  3. 3.
    Sarramegna V, Talmont F, Demange P, Milon A (2003) Heterologous expression of G-protein-coupled receptors: comparison of expression systems from the standpoint of large-scale production and purification. Cell Mol Life Sci 60:1529–1546PubMedCrossRefGoogle Scholar
  4. 4.
    McCusker EC, Bane SE, O’Malley MA, Robinson AS (2007) Heterologous GPCR expression: a bottleneck to obtaining crystal structures. Biotechnol Prog 23:540–547PubMedCrossRefGoogle Scholar
  5. 5.
    Tate CG, Grisshammer R (1996) Heterologous expression of G-protein-coupled receptors. Trends Biotechnol 14:426–430PubMedCrossRefGoogle Scholar
  6. 6.
    Ishihara G et al (2005) Expression of G protein coupled receptors in a cell-free translational system using detergents and thioredoxin-fusion vectors. Protein Expr Purif 41:27–37PubMedCrossRefGoogle Scholar
  7. 7.
    Klammt C et al (2007) Cell-free production of G protein-coupled receptors for functional and structural studies. J Struct Biol 158:482–493PubMedCrossRefGoogle Scholar
  8. 8.
    Corin K et al (2011) A robust and rapid method of producing soluble, stable, and functional G-protein coupled receptors. PLoS One 6(10):e23036PubMedCrossRefGoogle Scholar
  9. 9.
    Kaiser L et al (2008) Efficient cell-free production of olfactory receptors: detergent optimization, structure, and ligand binding analyses. Proc Natl Acad Sci 105(41):15726–15731PubMedCrossRefGoogle Scholar
  10. 10.
    Corin K et al (2011) Designer lipid-like peptides: a class of detergents for studying functional olfactory receptors using commercial cell-free systems. PLoS One 6(11):e25067PubMedCrossRefGoogle Scholar
  11. 11.
    Wang X et al (2011) Peptide surfactants for cell-free production of functional G protein-coupled receptors. Proc Natl Acad Sci USA 108(22):9049–9054PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  • Karolina Corin
    • 1
  • Brian Cook
    • 1
  • Shuguang Zhang
    • 1
  1. 1.Center for Bits and Atoms, Massachusetts Institute of TechnologyCambridgeUSA

Personalised recommendations