Structure and Mechanism of the Kinesin-3 Motor KIF1A

Kinesin-3 电机 KIF1A 的结构和机制

基本信息

批准号：
10735818
负责人：
Arne Gennerich
金额：
$ 62.17万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-01 至 2027-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10735818
关键词：
Affect Amino Acids Atrophic Axon Binding Biological Assay Brain Stem Cell Nucleus Cell division Cell physiology Complex Cryoelectron Microscopy Data Degenerative Disorder Dense Core Vesicle Disease Docking Dyes Elements Encephalopathies Engineering Etiology Exhibits Family Fluorescence Fluorescence Resonance Energy Transfer Frequencies Functional disorder Generations Goals Head Human In Vitro Individual Inherited Intellectual functioning disability Kinesin Label Link Measurement Mediator Microcephaly Microtubules Modernization Molecular Molecular Motors Motion Motor Mutagenesis Mutation N-terminal Neck Neurodegenerative Disorders Neurodevelopmental Disorder Neurons Nucleotides Optic Nerve Output Pattern Peripheral Nervous System Diseases Phenotype Physiological Plus End of the Microtubule Positioning Attribute Presynaptic Terminals Property Protein Engineering Proteins Resolution Series Severities Spastic Paraplegia Structural defect Structure Synaptic Vesicles System Testing Therapeutic Therapeutic Intervention Time Work autism spectrum disorder autonomic neuropathy cell motility cerebral atrophy de novo mutation developmental disease disease-causing mutation drug development falls human disease improved innovation insight laser tweezer member migration molecular targeted therapies mutant nervous system disorder neuron development novel nuclear power optic tweezer single molecule stem cells tool unnatural amino acids

项目摘要

Our goal is to define the structural and functional mechanism by which the kinesin-3 motor KIF1A generates force and moves along microtubules, and to define the structural basis of KIF1A-related human diseases. The microtubule (MT) transport system regulates essential eukaryotic activities, including neuronal cell division, neuronal migration and the transport of subcellular cargoes. KIF1A, a member of the kinesin-3 family of MT-associated motor proteins, is a key mediator of these activities as the major generator of MT plus-end-directed motility in neurons. KIF1A’s cargoes include nuclei in dividing brain stem cells and synaptic vesicle precursors and dense core vesicles in axon terminals. Not surprisingly, its dysfunction is implicated in a growing number of neurodevelopmental and neurodegenerative disorders referred to as KIF1A-associated neurological disorder (KAND). Unfortunately, these diseases remain poorly understood, in part because KIF1A’s molecular mechanism remains unclear. For example, while most mutations occur in KIF1A’s motor domain, high-resolution structures of the KIF1A-MT complex do not exist. In addition, it remains unknown why Kif1A is superprocessive but easily gives up under load. In this proposal, we will combine cryo-electron microscopy, single-molecule fluorescence, and optical tweezers-based force measurements with innovative protein engineering to determine why KIF1A is a weak but superprocessive motor, define high-resolution structures of the KIF1A-MT complex as a function of KIF1A’s mechanochemical cycle and determine the structural defects caused by disease mutations in KIF1A. These studies will provide a new understanding of KAND and elucidate molecular targets for therapeutic interventions.

我们的目标是定义驱动蛋白 3 马达 KIF1A 的结构和功能机制产生力并沿着微管移动，并定义 KIF1A 相关的结构基础人类疾病的微管（MT）运输系统调节重要的真核活动，包括神经元细胞分裂、神经元迁移和亚细胞货物的运输。 KIF1A 是 MT 相关运动蛋白的驱动蛋白 3 家族的成员，是这些活动是 KIF1A 神经元中 MT 加末端定向运动的主要产生者。货物包括分裂脑干细胞和突触小泡前体的核心和致密核心毫不奇怪，越来越多的轴突末端的囊泡与它的功能障碍有关。称为 KIF1A 相关的神经发育和神经退行性疾病不幸的是，人们对这些疾病的部分了解仍知之甚少。例如，因为 KIF1A 的分子机制仍不清楚，而大多数突变。由于 KIF1A-MT 复合物发生在 KIF1A 的运动域，因此不存在高分辨率结构。此外，目前还不清楚为什么 Kif1A 具有超处理能力，但在负载下很容易放弃。在这个提案中，我们将结合冷冻电子显微镜、单分子荧光和基于光镊的力测量与创新的蛋白质工程以确定原因 KIF1A 是一种弱但超处理的电机，定义了 KIF1A-MT 的高分辨率结构复杂作为 KIF1A 机械化学循环的函数并确定结构缺陷这些研究将为 KAND 提供新的认识。并阐明治疗干预的分子靶点。