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J. Biol. Chem., Vol. 282, Issue 20, 15284-15293, May 18, 2007

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Synergism of Accessory Factors in Functional Expression of Mammalian Odorant Receptors^*

Hanyi Zhuang¹ and Hiroaki Matsunami²

From the Department of Molecular Genetics and Microbiology, Department of Neurobiology, Duke University Medical Center, Durham, North Carolina 27710

The discovery of odorant receptors led to endeavors in matching them with their cognate ligands. Although it has been challenging to functionally express odorant receptors in heterologous cells, previous studies have linked efficient odorant receptor expression with N-terminal modifications and accessory proteins, including the receptor-transporting proteins (RTPs) and Ric8b. Here we have shown that a shorter form of RTP1, RTP1S, supports robust cell-surface and functional expression of representative odorant receptors. Using a combination of accessory proteins, including RTP1S, Ric8b, and G_olf, a diverse set of untagged odorant receptors were successfully expressed heterologously due to the synergistic effects among the various accessory proteins. Furthermore, the addition of an N-terminal rhodopsin tag to the odorant receptors, along with the same set of accessory proteins, exhibits an additional level of synergism, inducing enhanced odorant receptor responses to odorants and thus defining a more efficient heterologous expression system. We then showed that the presence or absence of different N-terminal tags has little effect on the ligand specificity of odorant receptors, although the amount of receptor expressed can play a role in the ligand response profile. The accuracy of the odorant receptor heterologous expression system involving tagged odorant receptors and various accessory proteins promises success in high throughput de-orphaning of mammalian odorant receptors.

Received for publication, January 16, 2007 , and in revised form, March 9, 2007.

^* This work was supported by Grant DC05782 from the National Institutes of Health (to H. M.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ Supported by the Ruth L. Kirschstein National Research Service Award from the National Institutes of Health (DC008480 [GenBank] ).

² To whom correspondence should be addressed: Dept. of Molecular Genetics and Microbiology, Duke University Medical Ctr., 264 CARL Bldg., Research Drive, Durham, NC 27710. Tel.: 919-684-2777; Fax: 919-681-9193; E-mail: hiroaki.matsunami{at}duke.edu.

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