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Originally published In Press as doi:10.1074/jbc.M203194200 on June 13, 2002

J. Biol. Chem., Vol. 277, Issue 35, 31459-31465, August 30, 2002
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Identification of a Novel Human Eicosanoid Receptor Coupled to Gi/o*

Takeshi HosoiDagger , Yutaka KoguchiDagger , Emiko Sugikawa, Aiko Chikada, Koji Ogawa, Naoki Tsuda, Naoki Suto, Shiho Tsunoda, Tomoyasu Taniguchi, and Tetsuo Ohnuki§

From the Discovery Research Laboratory, Tanabe Seiyaku Co. Ltd., 2-50 Kawagishi-2-chome, Toda-shi, Saitama 335-8505, Japan

We have conducted an in silico data base search for and cloned a novel G-protein-coupled receptor (GPCR) named TG1019. Dot and Northern blotting analyses showed that transcripts of the novel GPCR were expressed in various tissues except brain, and the expression was more intense in liver, kidney, peripheral leukocyte, lung, and spleen than in other tissues. By GTPgamma S binding assay using the TG1019-Galpha i1-protein fusion expressed in insect cells, eicosanoids, and polyunsaturated fatty acids such as 5-oxo-6E,8Z,11Z,14Z-eicosatetraenoic acid (5-oxo-ETE), 5(S)-hydroperoxy-6E,8Z, 11Z,14Z-eicosatetraenoic acid, and arachidonic acid were identified to exhibit agonistic activities against TG1019. 5-oxo-ETE was the most potent to enhance the specific binding by 6-fold at a maximum effect dose of submicromolar to micromolar order with an ED50 value of 5.7 nM. Conversely, polyunsaturated fatty acids such as docosahexaenoic acid and eicosapentaenoic acid showed antagonistic activities against TG1019. In Chinese hamster ovary cells transiently expressing TG1019, the forskolin-stimulated production of cAMP was inhibited up to ~70% by 5-oxo-ETE, with an IC50 value of 33 nM. This inhibition was sensitive to pretreatment of the cells with pertussis toxin.

* The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EBI Data Bank with accession number(s) AB083055.

Dagger Both authors contributed equally to this work.

§ To whom correspondence should be addressed. Tel.: 81-48-433-8065; Fax: 81-48-433-8159; E-mail: t-ohnuki@tanabe.co.jp.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.


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