Structural and Functional Studies of Rhodopsin and G-Protein Coupled Receptor Kinases

视紫红质和 G 蛋白偶联受体激酶的结构和功能研究

• 批准号: 10012941
• 负责人: Karsten Melcher
• 金额: $ 36.58万
• 依托单位: VAN ANDEL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-04-01 至 2021-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10012941
• 关键词: ARNT gene; Adopted; Agonist; Antibodies; Architecture; Arrestins; Binding; Binding Sites; Biochemical; Biological Assay; Biological Models; Cell Surface Receptors; Cells; Chemicals; Collaborations; Communication; Complex; Coupling; Cryoelectron Microscopy; Crystallization; Disease; Disulfides; Drug Targeting; Family; Future; G Protein-Coupled Receptor Signaling; G protein coupled receptor kinase; G-Protein-Coupled Receptors; GRK1 gene; GRK5 gene; GTP-Binding Proteins; Genetic Polymorphism; Goals; Heart Diseases; Homeostasis; Human; Knowledge; Ligands; Lipids; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Modeling; Molecular Conformation; Mutation; Nature; Negative Staining; Neurodegenerative Disorders; Pathway interactions; Phosphorylation; Phosphotransferases; Physiological; Process; Receptor Signaling; Resolution; Rhodopsin; Signal Transduction; Signaling Protein; Structure; Technology; Therapeutic; Time; base; biophysical techniques; crosslink; design; drug discovery; insight; interest; protein activation; receptor; receptor binding; screening; side effect; small molecule; success

PROJECT SUMMARY/ABSTRACT Humans have more than 800 different G protein-coupled receptors (GPCRs), which form the largest family of cell-surface receptors and account for ~30% of all therapeutic drug targets. Upon activation by agonists, GPCRs adopt distinct activated states that allow them to couple either to G proteins, to initiate G protein-mediated signaling, or to GPCR kinases (GRKs), which phosphorylate GPCRs to enable their interactions with arrestins to terminate G protein activation and initiate arrestin-mediated signaling. While physiological agonists initiate both G protein- and arrestin-mediated signaling, often only one of the two pathways is therapeutically beneficial, while the other pathway is responsible for unwanted side effects. Given the importance of GPCRs as drug targets, there is huge interest in developing “biased” GPCR agonist that signal predominantly through only one of the two pathways. However, the critical barrier for rationally developing biased agonist is the limited information on how GRKs interact with GPCRs and the complete absence of any high resolution GPCR/GRK structure. Crystal and cryo-EM structures of four GPCRs in complex with G proteins and of one GPCR in complex with an arrestin have provided an emerging understanding of the receptor conformations required for G protein and arrestin coupling, yet a mechanistic understanding of how GRKs bind and phosphorylate GPCRs have remained elusive due to the highly transient nature of this interaction. The objective of this proposal is to bridge this critical gap in knowledge by exploiting the interaction between rhodopsin and GRK1 as a paradigm to determine the mechanistic and structural basis of the targetable interaction between GPCRS and GRKs. SIGNIFICANCE: Completion of the proposed aims will reveal the first mechanistic basis of GRK/GPCR recognition and reveal the GPCR/GRK conformation that enables GRK binding and receptor phosphorylation. This information is of paramount importance for the rational design of therapeutic biased agonists for the treatment of a broad spectrum of diseases.

项目摘要/摘要 人类具有800多种不同的G蛋白偶联受体（GPCR），它们形成了 最大的细胞表面受体家族占所有治疗药物靶标的30％。 激动剂激活后，GPCR采用不同的激活状态，使他们能够搭配 要么G蛋白，要启动G蛋白介导的信号传导，或者启动GPCR激酶（GRKS），该信号（GRKS） 磷酸化GPCR可以使其与逮捕蛋白的相互作用终止G蛋白 激活并引发抑制蛋白介导的信号传导。当身体激动剂启动两个g 蛋白质和逮捕蛋白介导的信号传导，通常只有两种途径中只有一种 有益的，而另一条途径则负责有害的副作用。鉴于 GPCR作为药物靶标的重要性，对发展“有偏见” GPCR具有很大的兴趣 激动剂主要通过两种途径之一发出信号。但是，关键 合理发展的偏见激动剂的障碍是GRK相互作用的有限信息 GPCR和完全没有任何高分辨率GPCR/GRK结构。水晶 和与G蛋白和复合物中的一个GPCR复合物的四个GPCR的冷冻EM结构 逮捕蛋白提供了对接收器构象的新兴理解 G蛋白和阻止蛋白偶联所需 由于高度短暂的性质，磷酸化GPCR仍然难以捉摸 相互作用。该提议的目的是通过 利用视紫红质和grk1之间的相互作用作为范式来确定 GPCR和GRK之间可靶向相互作用的机械和结构基础。 意义：拟议目标的完成将揭示第一的机械基础 grk/gpcr识别并揭示GPCR/GRK构象，使GRK结合和 受体磷酸化。对于合理设计，此信息至关重要 治疗偏见的激动剂治疗多种疾病。