Structural analysis of the human LRRK2

人类 LRRK2 的结构分析

• 批准号: 10734733
• 负责人: Ji Sun
• 金额: $ 45.5万
• 依托单位: ST. JUDE CHILDREN'S RESEARCH HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-17 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10734733
• 关键词: AGFG1 gene; Accounting; Address; Affect; Aging; American; Architecture; Biochemical; Biological Assay; Biophysics; Cells; Cellular Assay; Cellular biology; Complex; Cryoelectron Microscopy; DNA Sequence Alteration; Data; Development; Disease; Family; Foundations; Genes; Genetic; Guanosine Triphosphate Phosphohydrolases; Histologic; Human; Idiopathic Parkinson Disease; Image; In Vitro; Knockout Mice; Knowledge; LRRK2 gene; Lead; Length; Lysosomes; Membrane; Molecular; Molecular Conformation; Monomeric GTP-Binding Proteins; Movement Disorders; Mutagenesis; Mutation; Neurodegenerative Disorders; Outcome; Parkinson Disease; Persons; Pharmacologic Substance; Phenotype; Phosphorylation; Phosphotransferases; Play; Process; Proteins; Research; Risk Factors; Role; Structure; Substrate Specificity; Techniques; Tertiary Protein Structure; Work; combat; design; drug development; drug discovery; experience; gain of function mutation; inhibitor; novel; novel strategies; novel therapeutics; particle; rab GTP-Binding Proteins; reconstitution; recruit; risk variant; sporadic Parkinson' s Disease; structural biology; trans-Golgi Network; vesicle transport

PROJECT SUMMARY Parkinson’s Disease (PD) is the second most common neurodegenerative disease, featured by movement disorders. PD affects nearly one million Americans and ten million people worldwide. While aging is the major risk factor, PD can also be caused by genetic mutations. More than ten PD-associated genes have been identified, and LRRK2 mutations are the most common cause of both familial and sporadic PD cases. LRRK2 gene encodes the leucine-rich repeat kinase 2 (LRRK2), a large multidomain protein with both kinase and GTPase activities; most of the LRRK2 disease mutations show increased kinase activity. Therefore, inhibiting LRRK2 kinase activity holds great potential for treating PD, and numerous pharmaceutical companies are making tremendous efforts to develop specific inhibitors. Our previous work has revealed the first cryo-EM structures of full-length human LRRK2, providing the atomic architecture of this 286-kDa kinase in an inactive conformation. Here we propose to further investigate the working mechanism of LRRK2 by combining cutting- edge structural biology techniques such as single-particle cryoEM with biochemical, biophysical and cell biology approaches. We hypothesize that understanding the structural basis of the LRRK2 recruitment, activation and substrate recognition could provide novel directions for the development of allosteric inhibitors. In this proposal, the working mechanism of LRRK2 will be dissected at atomic details by addressing the following fundamental questions. i) How do Rab GTPases recruit LRRK2? ii) What is the active conformation of LRRK2? and iii) how does LRRK2 specifically recognize its substrates? The successful outcome of this proposal will provide a molecular picture of the LRRK2 recruitment, activation and substrate recognition processes in atomic details and, therefore, could serve as the conceptual framework for designing allosteric LRRK2 inhibitors to treat PD.

项目概要 帕金森病 (PD) 是第二常见的神经退行性疾病，以运动为特征 PD 影响着近 100 万美国人和全球 1000 万人，而老龄化是其中的主要原因。 PD的危险因素也可能是由基因突变引起的，目前已发现十多个与PD相关的基因。 LRRK2 突变是家族性和散发性 PD 病例的最常见原因。 基因编码富含亮氨酸的重复激酶 2 (LRRK2)，这是一种大型多结构域蛋白，具有激酶和 GTPase 活性；大多数 LRRK2 疾病突变表现出激酶活性增加。 LRRK2 激酶活性在治疗 PD 方面具有巨大潜力，许多制药公司都在 我们付出了巨大的努力来开发特定的抑制剂，并揭示了第一个冷冻电镜。 全长人类 LRRK2 的结构，提供了这种 286-kDa 激酶在非活性状态下的原子结构 在这里，我们建议通过结合切割来进一步研究LRRK2的工作机制。 边缘结构生物学技术，例如结合生化、生物物理和细胞的单粒子冷冻电镜 我们追求了解 LRRK2 募集的结构基础， 激活和底物识别可以为变构的发展提供新的方向 在本提案中，LRRK2 的工作机制将通过原子细节进行剖析。 解决以下基本问题 i) Rab GTPase 如何招募 LRRK2？ LRRK2 的活性构象？以及 iii) LRRK2 如何特异性识别其底物？ 该提案的成功结果将提供 LRRK2 招募、激活和 原子细节中的基板识别过程，因此可以作为概念框架 设计变构 LRRK2 抑制剂来治疗 PD。