Signalling profiles of H3 relaxin, H2 relaxin and R3(B.23-27) R/I5 acting at the relaxin family peptide receptor 3 (RXFP3)
Signalling profiles of H3 relaxin, H2 relaxin and R3(B.23-27) R/I5 acting at the relaxin family peptide receptor 3 (RXFP3)
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DOI：
10.1111/bph.12623
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发表时间：
2014-06-01
影响因子：
7.3
通讯作者：
Summers, R. J.
中科院分区：
文献类型：
--
作者：
Kocan, M.;Sarwar, M.;Summers, R. J.
背景和目的放松蛋白家族肽受体 3 (RXFP3) 在处理感觉信息和进食的重要大脑区域中表达，表明它可能是抗焦虑和抗肥胖药物的靶点。我们检查了 H3 松弛素、偏向激动剂 H2 松弛素和拮抗剂 R3(B23-27)R/I5 对 RXFP3 信号传导的影响，以确定它们作为评估 RXFP3 生理作用的工具的适用性。实验方法使用报告基因测定、多重信号传导测定和使用 BRET 直接检查受体-G 蛋白和受体-抑制蛋白相互作用来确定 RXFP3 配体的信号传导谱。结果H2松弛素激活p38MAPK和ERK1/2的功效低于H3松弛素，但对JNK1/2磷酸化的功效相似。 p38MAPK、JNK1/2 或 ERK1/2 的 H2 或 H3 松弛素激活涉及百日咳毒素敏感 G 蛋白。 R3(B23-27)R/I5 阻断 H3 松弛素 AP-1 报告基因激活，但不阻断 H2 松弛素 AP-1 激活或 H3 松弛素 NF-B 激活。 R3(B23-27)R/I5 激活 SRE 报告基因，但不抑制 H2 或 H3 松弛素 SRE 激活。 R3(B23-27)R/I5 阻断 H3 松弛素刺激的 p38MAPK 和 ERK1/2 磷酸化，但是 p38MAPK 和 ERK1/2 信号传导的弱部分激动剂。 R3(B23-27)R/I5 激活的 p38MAPK 不依赖于 G 蛋白。 H3 松弛素激活的 RXFP3 与 G(i2)、G(i3)、G(oA) 和 G(oB) 相互作用，而 H2 松弛素或 R3(B23-27)R/I5 仅诱导与 G(i2) 或 G(oB) 相互作用。只有 H3 松弛素促进 RXFP3/-arrestin 相互作用，而这种相互作用被 R3(B23-27)R/I5 阻断。结论和意义了解作用于 RXFP3 的药物的信号传导特征对于开发针对该受体的疗法至关重要。
Background and PurposeRelaxin family peptide receptor 3 (RXFP3) is expressed in brain areas important for processing sensory information and feeding, suggesting that it may be a target for anti-anxiety and anti-obesity drugs. We examined the effects of H3 relaxin, the biased agonist H2 relaxin and the antagonist, R3(B23-27)R/I5, on RXFP3 signalling to establish their suitability as tools to assess the physiological roles of RXFP3.Experimental ApproachThe signalling profile of the RXFP3 ligands was determined using reporter gene assays, multiplexed signalling assays and direct examination of receptor-G protein and receptor--arrestin interactions using BRET.Key ResultsH2 relaxin activated p38MAPK and ERK1/2 with lower efficacy than H3 relaxin, but had similar efficacy for JNK1/2 phosphorylation. H2 or H3 relaxin activation of p38MAPK, JNK1/2 or ERK1/2 involved Pertussis toxin-sensitive G-proteins. R3(B23-27)R/I5 blocked H3 relaxin AP-1 reporter gene activation, but not H2 relaxin AP-1 activation or H3 relaxin NF-B activation. R3(B23-27)R/I5 activated the SRE reporter, but did not inhibit either H2 or H3 relaxin SRE activation. R3(B23-27)R/I5 blocked H3 relaxin-stimulated p38MAPK and ERK1/2 phosphorylation, but was a weak partial agonist for p38MAPK and ERK1/2 signalling. p38MAPK activation by R3(B23-27)R/I5 was G protein-independent. H3 relaxin-activated RXFP3 interacts with G(i2), G(i3), G(oA) and G(oB) whereas H2 relaxin or R3(B23-27)R/I5 induce interactions only with G(i2) or G(oB). Only H3 relaxin promoted RXFP3/-arrestin interactions that were blocked by R3(B23-27)R/I5.Conclusion and ImplicationsUnderstanding signalling profile of drugs acting at RXFP3 is essential for development of therapies targeting this receptor.