OPIOID RECEPTOR OLIGOMERIZATION: PREDICTION & VALIDATION

最近的研究发现，属于G蛋白偶联受体的阿片受体 (GPCR)家族，可在活细胞中以异构体形式存在，8-|x和6-K阿片受体复合体是 不同的功能信号单位，增加了阿片类药物研究领域的新的复杂性。 因此，关于阿片受体二聚体/低聚物复合体的结构和机理信息 对于了解鸦片类药物的作用机制具有重要意义。 在这项拨款申请中提出的工作的目标是识别分子决定因素 负责8-和|&gt；阿片受体(同构体和异构体)的齐聚 受体模型的结构背景，使用迭代结合计算和实验 接近。 由于没有关于阿片受体二聚体的详细结构数据，也没有证据表明人-阿片受体二聚体 与GPCRs的异位齐聚界面重合，我们开发了两种不同的计算方法 基于相关突变分析的方法，并确定了最有可能的同源和异源- 阿片受体亚型跨膜区之间的二聚化界面。推论来自 这些生物信息学工具和在初步研究中获得的初始分子模型构成了 将用于计算和实验相结合的策略的试点数据，如本文所述 批准应用，以探测、验证和改进8-和|I-阿片受体同源和 异构体。这些研究的一个重要目标是能够识别特定的突变 用于调节二聚化/寡聚化，从而影响受体功能 揭示依赖于二聚化和/或从二聚化而来的生理相关机制。 在此赠款申请中提出的计算和实验相结合的策略也是 有望为8-和|i-中亚型特有界面(S)的形成提供直接证据 阿片受体二聚体，或用于将这些受体排列成具有明显不同的高阶低聚物 对称或不对称界面，如最近提出的视紫红质低聚物。推论来自 这些研究有望为阿片受体的潜在机制提供新的见解。 功能，最终目的是帮助药物设计。

The recent discoveries that opioid receptors, which are members of the G-protein coupled receptor (GPCR) family, can exist as heteromers in live cells, and that 8-|x and 6-K opioid receptor complexes are distinct functional signaling units, have added a new dimension of complexity to the opioid research field. Structural and mechanistic information about opioid receptor dimeric/oligomeric complexes has therefore become of major importance for understanding the mechanisms of action of opiates. The goal of the work proposed in this grant application is to identify the molecular determinants responsible for the oligomerization of 8- and |>opioid receptors (both homo- and heteromers) in a structural context of receptor models, using an iterative combined computational and experimental approach. Since there is no detailed structural data about opioid receptor dimers, and no evidence that the homo- and hetero-oligomerization interfaces of GPCRs coincide, we developed two different computational approaches based on correlated mutation analysis, and identified the most likely homo- and hetero- dimerization interfaces between the transmembrane regions of opioid receptor subtypes. Inferences from these bioinformatics tools and initial molecular models obtained in the preliminary studies constitute the pilot data for the combined computational and experimental strategy that will serve, as described in this grant application, to probe, validate, and refine models of 8- and |i-opioid receptor homo- and heteromers. An important goal of these studies is to be able to identify specific mutations that can be used to modulate dimerization/oligomerization, and thus affect receptor function in a manner that will reveal physiologically relevant mechanisms that depend and/or ensue from dimerization. The combined computational and experimental strategy proposed in this grant application is also expected to provide direct evidence either for the formation of subtype-specific interface(s) in 8- and |i- opioid receptor dimers, or for the arrangement of these receptors into higher-order oligomers with distinct symmetrical or asymmetrical interfaces, as recently proposed for rhodopsin oligomers. Inferences from these studies are expected to provide new insights into the mechanisms underlying opioid-receptor function, with the ultimate goal of helping drug design.