Endosomes as a multifunctional hub to control GPCR function

项目摘要/摘要 GPCRs是最大的膜结合信号分子家族，是许多 目前在临床上使用的药物。传统上，gpcr被认为在细胞表面活跃，而不活跃。 同时在细胞内经历分子分离和贩运事件。最近的研究推翻了这一点 模特。GPCR在细胞内不是静止的。相反，现在已经知道GPCRs可以激活G蛋白 来自包括内小体在内的许多细胞内隔间的信号，以及这种细胞内信号 改变药物反应。虽然已经在努力将内体GPCR信号作为一种药物来利用 目标，关于内体的gpr分类以及这些贩运过程如何控制，人们知之甚少。 内体GPCR信号转导。这项提议的目标是解决围绕GPCR的知识差距。 对内体进行分类，并确定这些通路是如何控制内体信号的。在项目1中，我们测试 内体分选作为动力学计时器来控制内体的gpr信号的假说。 我们研究了一个典型的gpr，β2肾上腺素能受体，并使用了一种基因组合 工程学和蛋白质组学，以确定分选如何控制内体信号。在项目2中，我们重点关注 两个不同的GPCR，它们在内体发出信号，但缺乏任何一致的内体分选基序。我们 使用化学生物学、基因组学和蛋白质组学相结合的方法来确定 介导这些受体的内体分选。我们的研究试图揭示关于 保守了细胞生物学途径，同时推动了未来GPCR药物开发的新领域。

Project Summary/Abstract GPCRs are the largest family of membrane bound signaling molecules and, collectively, the target of many drugs currently used in the clinic. Classically, GPCRs were thought to be active at the cell surface and inactive while undergoing molecular sorting and trafficking events within the cell. Recent work has overturned this model. GPCRs are not quiescent inside of cells. Instead, it is now known that GPCRs can activate G protein signaling from many intracellular compartments including the endosome, and this intracellular signaling changes drug response. While efforts are already underway to harness endosomal GPCR signaling as a drug target, much is unknown about GPCR sorting at endosomes and how these trafficking processes control endosomal GPCR signaling. The goal of this proposal is to address the knowledge gap surrounding GPCR sorting at endosomes, and to determine how these pathways control endosomal signaling. In Project 1 we test the hypothesis that endosomal sorting functions as a kinetic timer to control GPCR signaling at endosomes. We examine a prototypical GPCR, the beta 2 adrenergic receptor, and the use a combination of genetic engineering and proteomics to determine how sorting controls endosomal signaling. In Project 2 we focus on two different GPCRs which signal at endosomes but lack any of the consensus endosomal sorting motifs. We use a combination of chemical biology, genomics, and proteomics to identify the proteins and pathways which mediate endosomal sorting of these receptors. Our studies seek to reveal fundamental lessons about conserved cell biological pathways while driving forward a new area for future GPCR drug development.