渭-opioid and 5-HT_1A receptors heterodimerize and show signalling crosstalk via G protein and MAP-kinase pathways

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• 作者：Didier Cussac ; Isabelle Rauly-Lestienne ; Peter Heusler ; Fré ; dé ; ric Finana ; Claudie Cathala ; Sophie Bernois ; Luc De Vries ; ^{luc.de.vries@pierre-fabre.com}
• 关键词：BRET ; Bioluminescence Resonance Energy Transfer ; FLAG ; Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys octapeptide epitope ; G伪 ; 伪 subunit of heterotrimeric G protein ; HA ; Hemagglutinin A epitope ; PTX ; pertussis toxin ; [35S]鈥怗TP纬S ; guanosine-5&prime ; -O-(3-[35S]thio)-
• 刊名：Cellular Signalling
• 出版年：2012
• 期刊代码：42_08986568
• 类别：bio
• 出版时间：August, 2012
• 卷：24
• 期：8
• 页码：1648-1657
• 文件大小：828 K

摘要

渭-opioid receptors have been shown to form heterodimers with several G protein coupled receptors involved in pain regulation such as 伪_2A-adrenergic and neurokinin 1 receptors. Because the 5鈥怘T_1A receptor is also involved in pain control, we investigated whether it can interact with the 渭-opioid receptor in cell lines. Using epitope-tagged 渭-opioid and 5鈥怘T_1A receptors, we show that both receptors can co-immunoprecipate when expressed in the same cells. This physical interaction was corroborated by a Bioluminescence Resonance Energy Transfer signal between the 渭-opioid receptor fused to Renilla luciferase and the 5鈥怘T_1A receptor fused to the Green Fluorescent Protein. Consistent with the presence of functional heterodimers, the 渭-opioid receptor activated a G伪_o protein covalently fused to the 5-HT_1A receptor in membrane preparations as well as a G伪₁₅ protein fused to the 5-HT_1A receptor in living cells. We demonstrate that both receptors can coexerce control of the ERK1/2 pathway: for example, 渭-opioid receptor-induced ERK1/2 phosphorylation was selectively desensitized by 5-HT_1A receptor activation. Although 5-HT_1A and 渭-opioid receptors were capable to internalize in response to their own activation, they were ineffective to induce the co-internalization of their partners. Thus, we show a functional heterodimerization of 渭-opioid and 5-HT_1A receptors in cell lines, a complex that might play a role in the control of pain in vivo. These results also support the potential therapeutic action of 5-HT_1A agonists against nociceptive processes.