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J. Biol. Chem., Vol. 280, Issue 26, 24816-24823, July 1, 2005

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Characterization of a Mouse Second Leukotriene B4 Receptor, mBLT2

BLT2-DEPENDENT ERK ACTIVATION AND CELL MIGRATION OF PRIMARY MOUSE KERATINOCYTES^*

Yoshiko Iizuka, Takehiko Yokomizo¶, Kan Terawaki, Mayumi Komine||, Kunihiko Tamaki||, and Takao Shimizu

From the Departments of Biochemistry and Molecular Biology and ^||Dermatology, Faculty of Medicine, The University of Tokyo and PRESTO of Japan Science and Technology Agency, Hongo 7-3-1, Bunkyo-ku, Tokyo 113-0033, Japan

Leukotriene B4 (LTB4) is a potent chemoattractant and activator for granulocytes and macrophages and is considered to be an inflammatory mediator. Two G-protein-coupled receptors for LTB4, BLT1 and BLT2, have been cloned from human and shown to be high and low affinity LTB4 receptors, respectively. To reveal the biological roles of BLT2 using mouse disease models, we cloned and characterized mouse BLT2. Chinese hamster ovary cells stably expressing mouse BLT2 exhibited specific binding to LTB4, LTB4-induced calcium mobilization, inhibition of adenylyl cyclase, and phosphorylation of extracellular signal-regulated kinase. We found that Compound A (4'-{[pentanoyl (phenyl) amino] methyl}-1, 1'-biphenyl-2-carboxylic acid) was a BLT2-selective agonist and induced Ca²⁺ mobilization and phosphorylation of extracellular signal-regulated kinase through BLT2, whereas it had no effect on BLT1. 12-epi LTB4 exhibited a partial agonistic activity against mBLT1 and mBLT2, whereas 6-trans-12-epi LTB4 did not. Northern blot analysis showed that mouse BLT2 is expressed highly in small intestine and skin in contrast to the ubiquitous expression of human BLT2. By in situ hybridization and the reverse transcriptase polymerase chain reaction, we demonstrated that mouse BLT2 is expressed in follicular and interfollicular keratinocytes. Compound A, LTB4, and 12-epi LTB4 all induced phosphorylation of extracellular signal-regulated kinase in primary mouse keratinocytes. Furthermore, Compound A and LTB4 induced chemotaxis in primary mouse keratinocytes. These data suggest the presence of functional BLT2 in primary keratinocytes.

Received for publication, November 24, 2004 , and in revised form, April 22, 2005.

^* This work was supported in part by Grants-in-aids from the Ministry of Education, Culture, Sports, Science and Technology of Japan. 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.

^¶ To whom correspondence should be addressed. Tel.: 81-3-5802-2925; Fax: 81-3-3813-8732; E-mail: yokomizo-tky{at}umin.ac.jp.

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