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Chemical Senses Advance Access published online on March 1, 2005
Chemical Senses, doi:10.1093/chemse/bji015
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© The Author 2005. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oupjournals.org
Accepted December 22, 2004

Article

Functional Characterization of Two Human Olfactory Receptors Expressed in the Baculovirus Sf9 Insect Cell System

Valéry Matarazzo 1 *, Olivier Clot-Faybesse 1 *, Brice Marcet 1, Gaëlle Guiraudie-Capraz 1 *, Boriana Atanasova 2, Gérard Devauchelle 3, Martine Cerutti 3, Patrick Etiévant 2, and Catherine Ronin 1*

1 UMR 6149 et GDR 2590 CNRS et Université de Provence, IFR du Cerveau, 31 Chemin J. Aiguier, F-13402 Marseille Cedex 20, France
2 Laboratoire de Recherches sur les Arômes, INRA, 17 rue Sully, BP 86510, 21065 Dijon Cedex, France
3 Station de Recherches de Pathologie Comparée, CNRS- INRA et GDR 2590, St Christol-Lez-Alès, France

* To whom correspondence should be addressed.
Catherine Ronin, E-mail: ronin{at}lnb.cnrs-mrs.fr


  Abstract

Olfactory receptors (ORs) are the largest member of the G-protein-coupled receptors which mediate early olfactory perception in discriminating among thousands of odorant molecules. Assigning odorous ligands to ORs is a prerequisite to gaining an understanding of the mechanisms of odorant recognition. The functional expression of ORs represents a critical step in addressing this issue. Due to limitations in heterologous expression, very few mammal ORs have been characterized, and so far only one is from human origin. Consequently, OR function still remains poorly understood, especially in humans, whose genome encodes a restricted chemosensory repertoire compared with most mammal species. In this study, we have designed cassette baculovirus vectors to coexpress human OR 17-209 or OR 17-210 with either G{alpha}olf or G{alpha}16 proteins in Sf9 cells. Each OR was found to be expressed at the cell surface and colocalized with both G{alpha} proteins. Using Ca2+ imaging, we showed that OR 17-209 and OR 17-210 proteins are activated by esters and ketones respectively. Odorant-induced calcium response was increased when ORs were coexpressed with G{alpha}16 protein ,whereas coexpression with G{alpha}olf abolished calcium signaling. This strategy has been found to overcome most of the limitations encountered when expressing an OR protein and has permitted odorant screening of functional ORs. Our approach could thus be of interest for further expression and ligand assignment of other orphan receptor proteins.

Keywords: baculovirus; calcium imaging; confocal microscopy; GPCR; insect cell system; olfactory receptor.

*Contributed equally to this paper and should be considered as first coauthors.


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