Endothelin-converting enzyme 2 differentially regulates opioid receptor activity
Endothelin-converting enzyme 2 differentially regulates opioid receptor activity
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DOI：
10.1111/bph.12833
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发表时间：
2015-01-01
影响因子：
7.3
通讯作者：
Devi, L. A.
中科院分区：
文献类型：
--
作者：
Gupta, A.;Fujita, W.;Devi, L. A.
背景和目的阿片受体的功能受受体内吞、再循环和/或降解等激活后事件的调节。虽然通常理解肽配体与受体共内吞，但相对较少的研究研究了内吞的肽和肽加工酶在调节受体功能中的作用。在这项研究中，我们专注于内皮素转换酶2（ECE 2），一个金属肽酶的脑啡肽酶家族的成员，表现出酸性pH最佳，定位于细胞内室，并选择性地处理神经肽，包括阿片肽在体外，实验方法ECE 2抑制对内吞肽水解的影响用薄层色谱法和使用ELISA或显微镜观察阿片受体转运。使用cAMP测定来测量ECE 2抑制对受体信号传导的影响，并且在体内，通过甩尾测定来测量对鞘内施用的阿片类药物诱导的抗伤害感受的影响。仅通过那些作为ECE 2底物的配体显著损害受体再循环和信号传导，这在异源细胞和内源性共刺激的细胞中均观察到。表达ECE 2受体。我们还发现，ECE 2抑制衰减的抗伤害感受介导的阿片肽ECE 2 substrates.Conclusions and ImplicationsThese结果表明，ECE 2，通过选择性地处理内源性阿片肽在内吞隔室，起着调节阿片受体activity.Linked Articles这篇文章是一个主题部分阿片类药物的一部分：新的途径功能选择性。要查看本节中的其他文章，请访问
Background and PurposeOpioid receptor function is modulated by post-activation events such as receptor endocytosis, recycling and/or degradation. While it is generally understood that the peptide ligand gets co-endocytosed with the receptor, relatively few studies have investigated the role of the endocytosed peptide and peptide processing enzymes in regulating receptor function. In this study, we focused on endothelin-converting enzyme 2 (ECE2), a member of the neprilysin family of metallopeptidases that exhibits an acidic pH optimum, localizes to an intracellular compartment and selectively processes neuropeptides including opioid peptides in vitro, and examined its role in modulating receptor recycling and resensitization.Experimental ApproachThe effect of ECE2 inhibition on hydrolysis of the endocytosed peptide was examined using thin-layer chromatography and on opioid receptor trafficking using either elisa or microscopy. The effect of ECE2 inhibition on receptor signalling was measured using a cAMP assay and, in vivo, on antinociception induced by intrathecally administered opioids by the tail-flick assay.Key ResultsThe highly selective ECE2 inhibitor, S136492, significantly impaired receptor recycling and signalling by only those ligands that are ECE2 substrates and this was seen both in heterologous cells and in cells endogenously co-expressing receptors with ECE2. We also found that ECE2 inhibition attenuated antinociception mediated only by opioid peptides that are ECE2 substrates.Conclusions and ImplicationsThese results suggest that ECE2, by selectively processing endogenous opioid peptides in the endocytic compartment, plays a role in modulating opioid receptor activity.Linked ArticlesThis article is part of a themed section on Opioids: New Pathways to Functional Selectivity. To view the other articles in this section visit