Discovering the mechanism of GPCR-mediated arrestin stimulation to enable effective drug therapies

项目摘要 导致G蛋白偶联受体(GPCRs)选择性刺激抑制素的药物，或 为了有选择地避免止痛药的刺激，承诺对 各种疾病，包括神经精神疾病、心血管疾病、肺病和新陈代谢疾病 精神错乱。尽管学术界和学术界都对gpr-arrestin的相互作用进行了深入的研究 制药业，尽管最近在结构生物学方面取得了巨大进展 GPCRs和逮捕，GPCRs刺激逮捕的机制仍然很差 明白了。同样，GPCRs可能实现支持或反对的选择性的手段 Arrestin信号仍不清楚。拟议中的研究将利用原子级分子。 动力学模拟以应对这些挑战，从而为 设计具有理想止痛药效果的功能性选择性gpr靶向药物。 目的1是确定arrestin的激活机制，准确地确定哪一种gpr- Arrestin相互作用表面驱动Arrestin活化和发现变构偶联 在arrestin区域之间，导致这些结构变化发生。剩下的 目的是确定gpcr构象(目标2)和gpcr的影响。 Arrestin结合和激活的磷酸化模式(目标3)。这将揭示一个gpcr如何 相对于G蛋白募集，可以支持或不支持arrestin募集和信号转导 发信号。它还将揭示gpr如何可能有利于特定的arrestin构象， 刺激Arrestin的一些下游效应，而不刺激其他效应。 拟议的研究将依赖于最先进的模拟方法，这些方法最近已经 能够确定GPCRs、G蛋白、转运蛋白和 其他蛋白质。它还将受益于与多名实验者的密切合作：结果 从结晶学、荧光光谱、核磁共振、电子顺磁共振和 细胞信号实验将结合起来指导和验证模拟。这项建议 意义重大，不仅因为它将阐明一个典型的生物信号传递过程，而且 还因为它将揭示GPCRs功能选择性的结构基础的一个关键部分。它 从而为合理设计更安全、更有效的药物提供基础 作用于GPCR，这是迄今为止最大的一类药物靶标。

Project Summary Medicines that cause G protein–coupled receptors (GPCRs) to selectively stimulate arrestins, or to selectively avoid stimulation of arrestins, promise more effective and safer treatments for a wide variety of diseases, including neuropsychiatric, cardiovascular, pulmonary and metabolic disorders. Despite intense study of GPCR–arrestin interactions in both academia and the pharmaceutical industry, and despite dramatic recent advances in the structural biology of GPCRs and arrestins, the mechanism by which GPCRs stimulate arrestins remains poorly understood. Likewise, the means by which GPCRs might achieve selectivity for or against arrestin signaling remains unclear. The proposed research will utilize atomic-level molecular dynamics simulations to address these challenges, thereby providing a foundation for the design of functionally selective GPCR-targeted drugs with desired effects on arrestins. Aim 1 is to determine the activation mechanism of arrestin, pinpointing which of the GPCR– arrestin interaction surfaces drives arrestin activation and discovering the allosteric coupling between regions of arrestin that causes these structural changes to take place. The remaining aims are to determine the effect of both GPCR conformation (Aim 2) and GPCR phosphorylation pattern (Aim 3) on arrestin binding and activation. This will reveal how a GPCR can favor or disfavor arrestin recruitment and signaling relative to G protein recruitment and signaling. It will also reveal how a GPCR can favor specific arrestin conformations, potentially stimulating some of arrestin’s downstream effects without stimulating others. The proposed research will rely on state-of-the-art simulation methods that have recently enabled the determination of functional mechanisms of GPCRs, G proteins, transporters, and other proteins. It will also benefit from close collaborations with multiple experimentalists: results from crystallography, fluorescence spectroscopy, NMR, electron paramagnetic resonance, and cell signaling experiments will combine to both guide and validate the simulations. This proposal is significant not only because it will illuminate a quintessential biological signaling process but also because it will reveal a key part of the structural basis for functional selectivity at GPCRs. It will thus provide a foundation for the rational design of safer and more effective medications acting at GPCRs, which are by far the largest class of drug targets.