Watching activation and signalling in individual GPCRs

观察单个 GPCR 的激活和信号传导

• 批准号: BB/G019738/1
• 负责人: Anthony Watts
• 金额: $ 83.9万
• 依托单位: University of Oxford
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2009
• 资助国家: 英国
• 起止时间: 2009 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FG019738%2F1
• 关键词: Watching; activation; signalling; individual; GPCRs

G-protein coupled receptors (GPCRs) represent one of the most important families of drug targets in pharmaceutical development. They are 7-transmembrane domain (7TMD) membrane receptors that control most neurological functions and comprise the largest family (~1%; >1,000 in number) of mammalian proteins. They are therefore at the focus of intense efforts in drug design and functional studies, not least because ~75% of drugs address only 5% of known GPCRs. We now (as of October 2007) have a structure of a ligand-binding Class A GPCR, the beta2-adrenergic receptor, to complement the 3D structures of rhodopsin. In the beta2-ADR structures, putative cholesterol binding sites and dimerisation interfaces were identified. Dimerisation of GPCRs may be fundamental to their signal activation mechanism, and lipid interactions crucial to ligand binding and/or the signalling process, both of which are areas of significant controversy and have only been partially addressed and not fully resolved. 3D structures are a very good start in such studies, but dynamics, the membrane involvement and kinetic mesurements are all essential in functional descriptions, even ahead of 3D structures. In particular, questions about ligand stoichiometry, interface interactions and lipid involvement in signalling are all important issues for understanding GPCR activation and signalling. The hurdle of obtaining a purified, reconstituted and functionally competent GPCR for such studies has been overcome for the chosen GPCR (neurotensin receptor 1). Studies now published by us (Harding, Biophys. J., 2008) demonstrate dimerisation in lipid bilayer of a specific composition. Here we will capitalize on this investment and use single molecule methods to dissect details about GPCR function using novel in vitro approaches for the first time on a GPCR.

g蛋白偶联受体（gpcr）是药物开发中最重要的药物靶点家族之一。它们是7-跨膜结构域（7TMD）膜受体，控制大多数神经功能，是哺乳动物蛋白中最大的家族（约占1%，数量约为1000个）。因此，它们是药物设计和功能研究的重点，尤其是因为约75%的药物只针对5%的已知gpcr。我们现在（截至2007年10月）有一个配体结合的a类GPCR的结构，β -肾上腺素能受体，以补充视紫红质的三维结构。在β 2- adr结构中，确定了假定的胆固醇结合位点和二聚化界面。GPCRs的二聚化可能是其信号激活机制的基础，而脂质相互作用对配体结合和/或信号传递过程至关重要，这两个领域都是具有重大争议的领域，目前只得到部分解决，尚未完全解决。3D结构在这类研究中是一个很好的开始，但是动力学、膜参与和动力学测量在功能描述中都是必不可少的，甚至在3D结构之前。特别是，配体化学计量学、界面相互作用和脂质参与信号传导的问题都是理解GPCR激活和信号传导的重要问题。对于选择的GPCR（神经紧张素受体1）来说，获得纯化的、重组的和功能有效的GPCR用于此类研究的障碍已经被克服。我们现在发表的研究(哈丁，生物物理学。J., 2008)证明在特定组成的脂质双分子层二聚化。在这里，我们将利用这一投资，并使用单分子方法解剖GPCR功能的细节，首次在GPCR上使用新颖的体外方法。

项目成果

Dynamic tuneable G protein-coupled receptor monomer-dimer populations.

• DOI: 10.1038/s41467-018-03727-6
• 发表时间: 2018-04-27
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Dijkman PM;Castell OK;Goddard AD;Munoz-Garcia JC;de Graaf C;Wallace MI;Watts A
• 通讯作者: Watts A