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: 9796468
• 负责人: John D McCorvy
• 金额: $ 34.84万
• 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/9796468
• 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

研究总结 G 蛋白偶联受体 (GPCR) 是涉及七种跨膜蛋白的重要超家族 细胞间通讯对于感知、运动和思维过程至关重要。 目前批准的药物疗法以 GPCR 为靶点，但会遭受与 GPCR 相关的“靶向”副作用 差异信号传导途径的参与被称为“功能选择性”或“偏向信号传导”。 偏向信号传递代表了一种有前途的方法，用于设计具有更好“on- 目标”概况，但导致信号偏差的结构水平机制仍然很差 理解偏向配体识别的结构知识的最新进展导致了这一点。 识别结合口袋基序，例如细胞外环 2 (EL2) 和跨膜 (TM) 7，非常重要 我的实验室使用基于结构的方法通过直接配体接合来“切换”偏向信号。 旨在使用各种化学生物学和生物物理方法来揭示内部的共同机制 控制偏向信号传导的结合口袋将结合结构引导的组合。 诱变、正构/变构偏向配体、G 蛋白功能和 β-arrestin 的动力学监测 使用荧光素酶和生物发光共振能量转移（BRET）技术进行招募，以及结构- “功能选择性”关系（SFSR）最终确定参与的关键 GPCR 结合基序 这种方法侧重于具有结构知识和 G 蛋白的关键受体。 通过这些研究，可以获得全面的机制了解。 对 GPCR 偏向信号的研究将引导研究人员开发新一代的优质疗法。 1