Endosomes as a multifunctional hub to control GPCR function

项目概要/摘要 GPCR是膜结合信号分子的最大家族，并且共同地，是许多靶分子的靶点。 目前临床上使用的药物。传统上，GPCR被认为在细胞表面是活性的，而在细胞表面是非活性的。 同时在细胞内经历分子分选和运输事件。最近的工作推翻了这一点 模型GPCR在细胞内不是静止的。相反，现在已知GPCR可以激活G蛋白 信号来自许多细胞内区室，包括内体，这种细胞内信号 改变药物反应。虽然已经在努力利用内体GPCR信号作为药物 靶向，很多是未知的GPCR分选在内涵体和如何这些运输过程控制 内体GPCR信号传导。本提案的目的是填补有关气相化学还原的知识空白 在核内体的分选，并确定这些途径如何控制核内体信号传导。在项目1中，我们测试 假设内体分选作为动力学计时器来控制内体的GPCR信号。 我们研究了一个原型GPCR，β 2肾上腺素能受体，并使用基因的组合， 工程和蛋白质组学来确定分选如何控制内体信号传导。在项目2中，我们关注 两种不同的GPCR，其在内体上发出信号，但缺乏任何共有的内体分选基序。我们 使用化学生物学，基因组学和蛋白质组学的组合来识别蛋白质和途径， 介导这些受体的内体分选。我们的研究试图揭示 保守的细胞生物学途径，同时推动未来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.