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.

项目总结 帕金森病(PD)是第二常见的神经退行性疾病，以运动为特征 精神错乱。帕金森病影响着近100万美国人和全球1000万人。虽然老龄化是主要的 危险因素中，帕金森病也可由基因突变引起。已有十多个帕金森氏症相关基因被 LRRK2突变是家族性和散发性帕金森病的最常见原因。LRRK2 LRRK2基因编码富含亮氨酸的重复蛋白2(LRRK2)，它是一种大的多结构域蛋白，既有激酶，也有 GTPase活性；大多数LRRK2疾病突变显示出激酶活性增加。因此，抑制 LRRK2激酶活性具有治疗帕金森病的巨大潜力，许多制药公司正在 做出巨大努力来开发特定的抑制剂。我们之前的工作揭示了第一个低温EM 全长的人LRRK2的结构，提供了这个286 kDa的非活性的 构象。在此，我们建议结合切割技术进一步研究LRRK2的作用机制。 生物化学、生物物理和细胞单粒子低温电子显微镜等边缘结构生物学技术 生物学走近了。我们假设，了解LRRK2招募的结构基础， 活化和底物识别为变构的发展提供了新的方向 抑制剂。在这份提案中，LRRK2的工作机制将通过以下方式进行原子细节剖析 解决以下基本问题。I)RAB GTP酶如何招募LRRK2？Ii)什么是 LRRK2活性构象？以及iii)LRRK2如何具体识别其底物？这个 这一提议的成功结果将提供LRRK2招募、激活和 原子细节中的底物识别过程，因此可以作为 设计变构LRRK2抑制剂治疗帕金森病。