Identification of farnesyl pyrophosphate and N-arachidonylglycine as endogenous ligands for GPR92
Identification of farnesyl pyrophosphate and N-arachidonylglycine as endogenous ligands for GPR92
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DOI：
10.1074/jbc.m708908200
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发表时间：
2008-07-25
影响因子：
4.8
通讯作者：
Seong, Jae Young
中科院分区：
文献类型：
--
作者：
Oh, Da Young;Yoon, Jung Min;Seong, Jae Young
使用信号通路特异性报告分析系统筛选了一系列作用于孤儿 G 蛋白偶联受体 GPR92 的小化合物。研究发现，包括法尼基焦磷酸 (FPP)、N-花生四烯酰甘氨酸 (NAG) 和溶血磷脂酸在内的脂质衍生分子可激活 GPR92。 FPP和溶血磷脂酸能够激活G(q/11)和G(s)介导的信号通路，而NAG仅激活G(q/11)介导的信号通路。计算机模拟模型结合 GPR92 的定点突变表明 GPR92 的 Thr97、Gly98、Phe101 和 Arg267 负责 GPR92 与 FPP 和 NAG 的相互作用。逆转录 PCR 分析显示，GPR92 mRNA 在背根神经节 (DRG) 中高表达，但在其他大脑区域中表达较弱。包括脾、胃、小肠和肾在内的外周组织也表达 GPR92 mRNA。免疫组织化学分析显示，在小鼠和人类 DRG 中，GPR92 很大程度上与 TRPV1 共定位，TRPV1 是一种对有害热做出反应的非特异性阳离子通道。 FPP 和 NAG 增加了培养的 DRG 神经元的细胞内 Ca2+ 水平。这些结果表明 FPP 和 NAG 通过激活 GPR92 在感觉神经系统中发挥作用。
A series of small compounds acting at the orphan G protein-coupled receptor GPR92 were screened using a signaling pathway-specific reporter assay system. Lipid-derived molecules including farnesyl pyrophosphate (FPP), N-arachidonylglycine (NAG), and lysophosphatidic acid were found to activate GPR92. FPP and lysophosphatidic acid were able to activate both G(q/11)- and G(s)-mediated signaling pathways, whereas NAG activated only the G(q/11)-mediated signaling pathway. Computer-simulated modeling combined with site-directed mutagenesis of GPR92 indicated that Thr97, Gly98, Phe101, and Arg267 of GPR92 are responsible for the interaction of GPR92 with FPP and NAG. Reverse transcription-PCR analysis revealed that GPR92 mRNA is highly expressed in the dorsal root ganglia (DRG) but faint in other brain regions. Peripheral tissues including, spleen, stomach, small intestine, and kidney also expressed GPR92 mRNA. Immunohistochemical analysis revealed that GPR92 is largely co-localized with TRPV1, a nonspecific cation channel that responds to noxious heat, in mouse and human DRG. FPP and NAG increased intracellular Ca2+ levels in cultured DRG neurons. These results suggest that FPP and NAG play a role in the sensory nervous system through activation of GPR92.