Detecting and directing molecular interactions that switch the functional state of G-protein coupled receptors

G蛋白偶联受体（GPCR）代表了几乎参与所有生理过程的最大一类膜蛋白（MP）。在分子水平上理解GPCR功能对现代生物学、医学和药理学提出了最重要的挑战之一。尽管这个蛋白质家族受到了强烈的关注，但对其作用的准确机制描述仍然没有解决。因此，了解定义GPCR功能状态的分子间和分子内相互作用是非常重要的。MP的单分子力谱（SMFS）是最近建立的方法，其允许检测和表征确定MP在其天然环境中的功能状态的相互作用。因此，该项目将利用这种现代生物物理方法来深入了解以前使用传统方法无法实现的GPCR功能。有了这些知识，我们将测试和优化配体样化合物来调节GPCR的功能。我们的方法将详细介绍天然状态下膜嵌入GPCR内的分子相互作用，这些受体的病理状态是如何实现的，以及突变GPCR如何被配体拯救以显示天然状态。

G protein-coupled receptors (GPCRs) represent the largest class of membrane proteins (MPs) that are involved in virtually every physiological process. Understanding GPCR function at a molecular level poses one of the most important challenges to modern biology, medicine, and pharmacology. Despite the intense focus this family of proteins has received, an accurate mechanistic description of their action remains unsolved. Thus, it is of great importance to understand the inter- and intramolecular interactions that define the functional state of GPCRs. Single-molecule force spectroscopy (SMFS) of MPs is a recently established method that allows for the detection and characterization of the interactions that determine the functional state of MPs in their native environment. This project will therefore utilize this modern biophysical approach to gain insight into the function of GPCRs that have been previously unattainable using traditional methods. With this knowledge we will test and optimize ligand-like compounds to modulate function of GPCRs. Our approach will detail molecular interactions within the membrane embedded GPCR in the native state, how the pathological state of these receptors is attained, and how mutant GPCRs can be rescued by a ligand to display the native state.