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.

项目摘要 帕金森病（Parkinson's Disease，PD）是第二常见的神经退行性疾病，以运动障碍为特征 紊乱PD影响着近100万美国人和全世界1000万人。而衰老是主要的 风险因素，PD也可以由基因突变引起。十多个PD相关基因已被发现。 LRRK 2突变是家族性和散发性PD病例的最常见原因。LRRK2 基因编码富含亮氨酸重复激酶2（LRRK 2），这是一种既具有激酶又具有蛋白质结构域的大型多结构域蛋白质。 GT3活性;大多数LRRK 2疾病突变显示激酶活性增加。因此，抑制 LRRK 2激酶活性具有治疗PD的巨大潜力，并且许多制药公司都在开发LRRK 2激酶。 在开发特异性抑制剂方面做出了巨大努力。我们之前的工作揭示了第一个冷冻电镜 全长人LRRK 2的结构，提供了这种286-kDa激酶的原子结构， 构象在此，我们建议进一步研究LRRK 2的工作机制，结合切割- 边缘结构生物学技术，如单粒子cryoEM与生物化学，生物物理和细胞 生物学方法。我们假设，了解LRRK 2募集的结构基础， 激活和底物识别可能为开发变构药物提供新的方向。 抑制剂的在本提案中，LRRK 2的工作机制将在原子细节上进行剖析， 解决以下基本问题。i）Rab GTP酶如何招募LRRK 2？（二）什么是 LRRK 2的活性构象？以及iii）LRRK 2如何特异性地识别其底物？的 该提议的成功结果将提供LRRK 2募集、激活和 衬底识别过程中的原子细节，因此，可以作为概念框架， 设计变构LRRK 2抑制剂以治疗PD。