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

发现 GPCR 介导的抑制蛋白刺激机制以实现有效的药物治疗

• 批准号: 10092188
• 负责人: Ron Dror
• 金额: $ 30.94万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-04-01 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10092188
• 关键词: Academia; Address; Adopted; Affect; Alzheimer' s Disease; Arrestins; Binding; Binding Proteins; Biological; Biophysics; C-terminal; Cardiovascular Diseases; Cardiovascular system; Collaborations; Complex; Computers; Coupling; Crystallization; Crystallography; Dangerousness; Dependence; Development; Diabetes Mellitus; Disease; Dissociation; Drug Industry; Drug Receptors; Drug Targeting; Electron Spin Resonance Spectroscopy; Electrons; Event; Family; Fluorescence Spectroscopy; Foundations; G-Protein-Coupled Receptors; GTP-Binding Protein alpha Subunits, Gs; GTP-Binding Proteins; Goals; Inflammation; Lead; Ligands; Lung; Lung diseases; Malignant Neoplasms; Mediating; Medicine; Metabolic Diseases; Methods; Molecular; Molecular Conformation; Obesity; Pathway interactions; Pattern; Pharmaceutical Preparations; Pharmacotherapy; Phosphorylation; Process; Property; Proteins; Publications; Research; Research Personnel; Resolution; Shapes; Signal Pathway; Signal Transduction; Structure; Surface; Tail; Techniques; Work; base; computer studies; design; drug discovery; effective therapy; experimental group; experimental study; molecular dynamics; neuropsychiatric disorder; novel therapeutics; protein activation; receptor; receptor binding; receptor coupling; recruit; scaffold; side effect; simulation; single-molecule FRET; structural biology; trafficking

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.

项目摘要 导致G蛋白偶联受体（GPCR）选择性刺激抑制蛋白的药物，或 选择性地避免抑制蛋白的刺激，承诺更有效和更安全的治疗， 多种疾病，包括神经精神、心血管、肺和代谢疾病 紊乱尽管在学术界和临床上对GPCR-抑制蛋白相互作用进行了大量的研究， 制药工业，尽管最近在结构生物学方面取得了巨大进展， GPCR和抑制蛋白，GPCR刺激抑制蛋白机制仍然很差 明白同样，GPCR可能实现选择性支持或反对 抑制蛋白信号传导仍然不清楚。拟议的研究将利用原子级分子 动态模拟来应对这些挑战，从而为 设计对抑制蛋白具有期望效果的功能选择性GPCR靶向药物。 目的1是确定arrestin的激活机制，确定GPCR- 抑制蛋白相互作用表面驱动抑制蛋白活化并发现变构偶联 引起这些结构变化的区域之间。其余 目的是确定GPCR构象（Aim 2）和GPCR 磷酸化模式（Aim 3）对抑制蛋白结合和活化的影响。这将揭示GPCR 相对于G蛋白募集，可以有利于或不利于抑制蛋白募集和信号传导， 发信号。它还将揭示GPCR如何有利于特定的抑制蛋白构象， 刺激了arrestin的一些下游效应，而没有刺激其他效应。 拟议中的研究将依赖于最先进的模拟方法， 能够确定GPCR、G蛋白、转运蛋白和 其他蛋白质。它还将受益于与多个实验者的密切合作：结果 晶体学、荧光光谱、NMR、电子顺磁共振， 细胞信号实验将结合联合收割机来指导和验证模拟。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导过程， 还因为它将揭示GPCR的功能选择性的结构基础的关键部分。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点。