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蛋白组合受体（GPCR），它们形成了 最大的细胞表面受体家族，并说明 激动剂激活后，GPCR采用不同的激活状态 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可以使与逮捕蛋白的相互作用与终末G蛋白相互作用 激活和启动逮捕蛋白介导的信号传导。 蛋白质和阻止蛋白介导的信号传导，通常只有一种治疗方法 有益的，而另一条途径则负责有害的副作用。 GPCR作为药物靶标的重要性，对发展“有偏见” GPCR的极大兴趣 激动剂主要通过两个途径之一 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和任何高分辨率GPCR/GRK结构的复合物。 和与G蛋白的复合物中的四个GPCR的冷冻EM结构和一个复合物中的一个GPCR 有被捕者已经证明了对受体信心的新兴理解 G蛋白和阻止蛋白偶联所需 并且磷酸化GPCR仍然难以捉摸 相互作用。 利用视紫红质和grk1之间的相互作用，高达多一点，只能多得多，只能多得多，只能像多样性的程度高达一定程度的程 GPCR和GRK之间可靶向相互作用的机械和结构基础。 意义：支撑目的的压缩将是第一个机械基础 GRK/GPCR识别并揭示GPCR/GRK Conforation，使GRK结合Anding 受体磷酸化。 治疗性偏见的激动剂，以涉及各种各样的疾病。