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Papers In Press, published online ahead of print June 13, 2002
Discovery Research Laboratory, Tanabe Seiyaku Company, Ltd., Toda-shi, Saitama 335-8505
Corresponding Author: t-ohnuki{at}tanabe.co.jp
We have conducted an in silico database 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 GTPgS binding assay using the TG1019-Gia1-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 of the basal binding, 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 CHO cells transiently expressing TG1019, the forskolin-stimulated production of cAMP was inhibited up to ca. 70 % by 5-oxo-ETE, with an IC50 value = 33 nM. This inhibition was sensitive to pretreatment of the cells with pertussis toxin.
J. Biol. Chem, 10.1074/jbc.M203194200
Submitted on April 3, 2002
Revised on June 10, 2002
Accepted on June 13, 2002
Identification of a novel eicosanoid receptor coupled to Gi/o
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