Identification of N-arachidonylglycine as the endogenous ligand for orphan G-protein-coupled receptor GPR18
Identification of N-arachidonylglycine as the endogenous ligand for orphan G-protein-coupled receptor GPR18
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DOI：
10.1016/j.bbrc.2006.06.175
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发表时间：
2006-09-01
影响因子：
3.1
通讯作者：
Yasukawa, Masaki
中科院分区：
文献类型：
--
作者：
Kohno, Masashi;Hasegawa, Hitoshi;Yasukawa, Masaki
根据人趋化因子受体保守区设计的引物，在对HUT 102细胞进行简并寡核苷酸聚合酶链式反应分析的基础上，克隆了一个孤儿G蛋白偶联受体GPR18。GPR18在淋巴系细胞系中显著表达，而在非淋巴系造血系细胞系中不表达。此外，GPR18基因在外周血淋巴细胞亚群(CD_4(+)、CD_4(+)、CD_(45)RA(+)、CD_4(+)、CD_(45)RO(+)、CD_8(+)、CD_(19)(+))中的表达高于单核细胞和淋巴样细胞系，植物血凝素刺激后GPR18基因的表达增强。通过脂质文库筛选，N-花生四烯基甘氨酸(NAGLY)诱导GPR18转染组细胞内钙离子浓度升高，明显高于模型转染组。在转GPR18的CHO细胞中，nAGLY还以百日咳毒素敏感的方式抑制Forsklin诱导的cAMP的产生。这是第一次证明nAGLY是GPR18的天然配体。(C)2006 Elsevier Inc.保留所有权利。
An orphan G-protein-coupled receptor, GPR18, was cloned on the basis of degenerate-oligonucleotide PCR analysis of HUT 102 cells using primers designed from the conservative regions of the human chemokine receptor. GPR18 was expressed significantly in lymphoid cell lines, but not in non-lymphoid hematopoietic cell lines. Moreover, the expression of the GPR18 gene was higher in peripheral lymphocyte subsets (CD4(+), CD4(+)CD45RA(+), CD4(+)CD45RO(+), CD8(+), and CD19(+)) than in monocytes and lymphoid cell lines, and was increased after stimulation with phytohemagglutinin. By screening using a lipid library, N-arachidonylglycine (NAGly) induced an increase in intracellular Ca2+ concentration in GPR18-transfected cells, which was significantly greater than that in mock-transfected cells. NAGly also inhibited forskolin-induced cAMP production in a pertussis toxin-sensitive manner in the GPR18-transfected CHO cells. This is the first study to demonstrate that NAGly is a natural ligand for GPR18. (c) 2006 Elsevier Inc. All rights reserved.