Assay to Screen Ligands:Activate Opioid Receptor (RMI)

G蛋白偶联受体构成了介导细胞凋亡的细胞表面受体的最大家族。 对各种刺激的生理反应。这些受体的特征是七个 跨膜结构域，并与异源三聚体G蛋白相互作用以抑制细胞信号。用于 多年来，我们的研究一直集中在了解调节阿片类药物的分子机制 受体;这些受体通过与经典的阿片类药物如吗啡结合而被激活。阿片受体 功能受到多种机制的调节;我们重点关注受体二聚化作为一种新的机制 调节阿片受体功能的机制。我们已经表征了异二聚体的性质 阿片受体类型（i.e.mu和δ阿片受体）之间的关系，使用生物化学，生物物理和 药理学技术我们已经发现，μ受体介导的细胞内信号转导和镇痛作用， 小鼠可以通过δ受体拮抗剂增强。这些结果表明， μ和δ受体导致信号传导程度的调节。为了解决生理问题 μ-δ异二聚化的结果，选择性激活或抑制μ-δ的配体 需要异二聚体（不影响μ或δ受体的活性）。在本申请中，我们 提出开发高通量筛选（HTS）测定以筛选选择性结合并 激活μ-δ异二聚体。这些研究将使HTS筛选和识别小 这些分子将是探测μ-δ异源二聚体在体内的作用的关键工具。

G-protein coupled receptors constitute the single largest family of cell surface receptors that mediate physiological responses to a wide variety of stimuli.These receptors are characterized by seven transmembrane domains and interact with heterotrimeric G-proteins to transduce cellular signals. For a number of years our studies have focused on understanding the molecular mechanisms that modulate opioid receptors; these receptors are activated by binding to classic opiates such as morphine. Opioid receptor function is modulated by multiple mechanisms; we have focused on receptor dimefization as a novel mechanism to regulate opioid receptor function. We have characterized the properties of heterodimers between opioid receptor types (i.e.mu and delta opioid receptors) using biochemical, biophysical and pharmacological techniques. We have found that the mu receptor-mediated signalingin cells and analgesia in mice can be enhanced by a delta receptor antagonist. These results suggest that heterodimerization between mu and delta receptors leads modulation of the extent of signaling. In order to address the physiological consequences of mu-delta heterodimerization, ligands that selectively activate or inactivate mu-delta heterodimers (without affecting the activities of mu or delta receptors) are needed. In this application we propose to develop a high throughput screening (HTS) assay to screen for ligands that selectively bind and activate mu-delta heterodimers. These studies will enable HTS screening and identification of small molecules that would be critical tools to probe the role of mu-delta heterodimers in vivo.