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

G蛋白偶联受体（gpcr）代表了最大的一类膜蛋白（MPs），几乎参与了每一个生理过程。在分子水平上理解GPCR的功能是现代生物学、医学和药理学面临的最重要挑战之一。尽管这一蛋白质家族受到了广泛关注，但对其作用的准确机制描述仍未得到解决。因此，了解分子间和分子内的相互作用是非常重要的，这些相互作用定义了gpcr的功能状态。MPs的单分子力谱（SMFS）是最近建立的一种方法，可以检测和表征决定MPs在其原生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学方法来深入了解gpcr的功能，这是以前使用传统方法无法实现的。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在天然状态下膜内嵌入GPCR的分子相互作用，如何获得这些受体的病理状态，以及如何通过配体拯救突变GPCR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.