CryoEM structure of cannabinoid receptor CB2 with a biased ligands

具有偏向配体的大麻素受体 CB2 的冷冻电镜结构

• 批准号: 2745686
• 金额: --
• 依托单位: University of Nottingham
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2745686
• 关键词: CryoEM; structure; cannabinoid; receptor; CB2

G protein-coupled receptors (GPCRs) are transmit signals between the cellular environment and its interior. Due to their key role in cellular signalling, they are the target of a third of marketed drugs. Binding of ligands with agonist activity stabilises particular conformations of a GPCR, which result in the activation of G proteins and arrestin-mediated signalling pathways. It is now widely recognized that "biased" therapeutics, which activate only a subset of specific signalling pathways, offer significant advancement over existing drugs with full agonist or antagonist activity due to higher specificity of action and lower side effects. Significant efforts are now being directed to develop such drugs. However, due to the promiscuity of the GPCRs and complexity of the signalling pathways, it is often difficult to attribute the observed effects of candidate biased ligands to one particular mode of signalling. Cannabinoid receptor CB2 is a valuable drug target for treating inflammatory diseases, pain, osteoporosis, atherosclerosis and brain tumours. We recently identified several CB2 ligands that show strong biased signalling profile. Complementary mutagenesis results suggested that the ligand binding site contains "hot-spots" that mediate signalling induced by the ligand binding, and are likely to affect biased signalling. Our main hypothesise is that CB2 ligands exert their action and signalling bias via specific interactions in the ligand binding pocket. To test this hypothesis, we need structural information of how biased ligands bind the receptor. The project will focus on obtaining a CryoEM structure of CB2 receptor in complex with several biased ligands using CryoEM. The student will learn mammalian cell culture, membrane protein biochemistry and single-particle CryoEM techniques. These skills will be complemented with training pharmacological assays to characterise pharmacology of novel ligands inspired by the structural data. The results of the project will establish a link between the structural changes induced in receptors to their signalling properties and would promote the development of the next generation of "biased" therapeutics.The student will be embedded in the Centre of Membrane Proteins and Receptors (COMPARE), a unique collaboration between the Universities of Birmingham and Nottingham, a centre of GPCR excellence in Europe and worldwide. It offers a vibrant community of over 40 PhD students and post-docs to develop research and presentation skills and an academic network. Additionally, they will work at RCH on CryoEM structure determination and further expanding their academic network.

G蛋白偶联受体（GPCR）是细胞环境及其内部之间的发射信号。由于它们在细胞信号中的关键作用，它们是销售药物的三分之一的目标。具有激动剂活性的配体的结合稳定了GPCR的特定构象，从而导致G蛋白的激活和阻滞蛋白介导的信号传导途径。现在，人们广泛认识到，仅激活特定信号通路的一部分“有偏见”的治疗剂，由于动作的特异性和较低的副作用，因此对具有全部激动剂或拮抗剂活性的现有药物提供了显着进步。现在正在指示开发此类药物的重大努力。但是，由于GPCR的滥交和信号通路的复杂性，通常很难将观察到的候选配体的效应归因于一种特定的信号传导方式。大麻素受体CB2是治疗炎症性疾病，疼痛，骨质疏松症，动脉粥样硬化和脑肿瘤的宝贵药物。我们最近确定了几种CB2配体，这些配体显示出强烈的信号传导曲线。补充诱变结果表明，配体结合位点包含“热点”，可介导由配体结合引起的信号传导，并可能影响偏置信号传导。我们的主要假设是CB2配体通过配体结合袋中的特定相互作用发挥其作用和信号偏差。为了检验这一假设，我们需要有关偏置配体如何结合受体的结构信息。该项目将专注于使用冷冻物的几种有偏的配体在复合物中获得CB2受体的冷冻结构。学生将学习哺乳动物细胞培养，膜蛋白生物化学和单粒子冷冻技术。这些技能将与培训药理学测定法相辅相成，以表征受结构数据启发的新型配体的药理学。 The results of the project will establish a link between the structural changes induced in receptors to their signalling properties and would promote the development of the next generation of "biased" therapeutics.The student will be embedded in the Centre of Membrane Proteins and Receptors (COMPARE), a unique collaboration between the Universities of Birmingham and Nottingham, a centre of GPCR excellence in Europe and worldwide.它提供了一个充满活力的社区，由40多名博士生和博士后，以发展研究和演示技巧和学术网络。此外，他们还将在RCH上进行冷冻结构的确定，并进一步扩大其学术网络。