Molecular Mechanisms of G protein-coupled Receptor Biased Signaling

G 蛋白偶联受体偏向信号传导的分子机制

• 批准号: 10164807
• 负责人: John D McCorvy
• 金额: $ 38.5万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10164807
• 关键词: Binding; Biological Assay; Biology; Cell Communication; Cells; Chemicals; Drug Targeting; Energy Transfer; Exhibits; G-Protein-Coupled Receptors; GTP-Binding Proteins; Generations; Integral Membrane Protein; Kinetics; Knowledge; Laboratories; Lead; Ligands; Luciferases; Molecular; Monitor; Movement; Mutagenesis; Pathway interactions; Pharmaceutical Preparations; Process; Research; Research Personnel; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Structure; Techniques; Therapeutic; base; beta-arrestin; biophysical techniques; design; extracellular; preference; protein B; protein function; receptor; receptor binding; recruit; side effect; targeted treatment

Research Summary G protein-coupled receptors (GPCRs) are an important superfamily of seven transmembrane proteins involved in cell-to-cell communication essential for sensing, movement, and thought processes. A significant portion of current approved drug therapies target GPCRs, but suffer from “on-target” side-effects related to the engagement of differential signaling pathways known as “functional selectivity” or “biased signaling.” Exploiting biased signaling represents a promising approach toward designing pathway-selective drugs with better “on- target” profiles, but the mechanisms at the structural level that lead to biased signaling are still poorly understood. Recent advancement in structural knowledge of biased ligand recognition has led to the identification of binding pocket motifs, such as extracellular loop 2 (EL2) and transmembrane (TM) 7, important for `switching' biased signaling via direct ligand engagement. Using a structure-based approach, my laboratory aims to use a variety of chemical biology and biophysical approaches to uncover common mechanisms within the binding pocket that govern biased signaling. Strategies will incorporate a combination of structure-guided mutagenesis, orthosteric/allosteric biased ligands, kinetic monitoring of G protein function and β-arrestin recruitment using luciferase and bioluminescent resonance energy transfer (BRET) techniques, and structure- `functional selectivity' relationships (SFSRs) to ultimately to pin-point key GPCR binding motifs involved in effector switching. This approach focuses on key receptors where there is structural knowledge and G protein and β-arrestin-biased ligands available. Through these studies, a comprehensive mechanistic understanding into GPCR biased signaling will guide researchers toward a new generation of superior therapeutics. 1

研究摘要 G蛋白偶联受体（GPCR）是涉及七个跨膜蛋白的重要超家族 在感测，运动和思考过程至关重要的细胞间通信中。大部分 当前已批准的药物疗法针对GPCR，但遭受了与 差异信号通路的参与被称为“功能选择性”或“偏置信号传导”。利用 偏见的信号传导代表了设计途径选择性药物的一种有希望的方法 目标”曲线，但是导致信号传导的结构水平的机制仍然很差 理解。偏向配体认识的结构知识的最新进步已导致 鉴定结合袖珍主题，例如细胞外环2（EL2）和跨膜（TM）7，重要 用于“切换”通过直接配体参与的偏置信号传导。使用基于结构的方法，我的实验室 旨在使用各种化学生物学和生物物理方法来发现内部的共同机制 结合信号传导的结合袋。策略将结合结构引导的组合 诱变，直角/变构偏配体，G蛋白功能的动力学监测和β-arrestin 使用荧光素酶和生物发光共振能量转移（BRET）技术募集，结构 - “功能选择性”关系（SFSR）最终到针对参与的PIN点键GPCR结合基序 效应器切换。这种方法着重于具有结构知识和G蛋白的关键受体 可用和β-art蛋白偏置配体。通过这些研究，一种全面的机械理解 进入GPCR偏见的信号传导将指导研究人员进行新一代的上级治疗。 1