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电动机KIF1A的结构和功能机制产生力并沿着微管移动，并定义与KIF1A相关的结构基础人类疾病。微管（MT）运输系统调节基本真核活动，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，是这些活动是神经元中MT Plus指导运动的主要发生者。 kif1a的货物包括在脑干细胞和突触囊泡前体和致密核心的核心轴突末端的囊泡。毫不奇怪，其功能障碍在越来越多的神经发育和神经退行性疾病称为KIF1A相关神经系统障碍（KAND）。不幸的是，这些疾病的理解程度很高，部分原因是因为KIF1A的分子机制尚不清楚。例如，大多数突变发生在KIF1A的运动域中，不存在KIF1A-MT复合物的高分辨率结构。此外，为什么KIF1A是超级过程，但很容易在负载下放弃。该建议，我们将结合冷冻电子显微镜，单分子荧光和具有创新蛋白工程的基于光学镊子的力量测量，以确定为什么 KIF1A是弱但超级过程的电动机，定义了KIF1A-MT的高分辨率结构复合物是KIF1A机械化学循环的函数，并确定结构缺陷由KIF1A中的疾病突变引起。这些研究将提供对Kand的新理解并阐明用于治疗干预措施的分子靶标。