Structural characterization of APP family proteins

APP 家族蛋白的结构表征

• 批准号: 10648792
• 负责人: Vasileios I Petrou
• 金额: $ 15.7万
• 依托单位: RUTGERS BIOMEDICAL AND HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10648792
• 关键词: Adult; Affect; Alzheimer' s Disease; American; Amyloid; Amyloid beta-Protein; Amyloid beta-Protein Precursor; Architecture; Behavior; Biochemical; Biological; Biological Process; Birth; Brain; C-terminal; Cell surface; Communication; Complement; Cryoelectron Microscopy; Dementia; Deposition; Detergents; Disease; Drug Design; Early Onset Familial Alzheimer' s Disease; Elements; Exhibits; Family member; Functional disorder; Future; Gene Family; Gene Proteins; Goals; Homeostasis; Individual; Inherited; Intracellular Transport; Investigation; Knock-out; Learning; Length; Link; Location; Maintenance; Mammalian Cell; Membrane; Membrane Glycoproteins; Membrane Proteins; Methods; Microscope; Molecular; Molecular Sieve Chromatography; Mutation; N-Methyl-D-Aspartate Receptors; Nerve Degeneration; Nervous System; Neurites; Neurodegenerative Disorders; Neurofibrillary Tangles; Neurons; Nicotinic Receptors; Orthologous Gene; Pathogenesis; Pathologic; Pathway interactions; Pattern; Peptides; Performance; Physiological; Play; Positioning Attribute; Preparation; Procedures; Process; Program Description; Proliferating; Property; Protein Biochemistry; Protein Family; Proteins; Recycling; Regulation; Reporting; Research; Resolution; Resources; Role; Sampling; Senile Plaques; Shapes; Signal Transduction; Structure; Synapses; Synaptic Vesicles; Synaptic plasticity; System; Techniques; Vertebral column; X-Ray Crystallography; abeta accumulation; beta secretase; cerebral atrophy; cryogenics; density; early onset; experience; extracellular; familial Alzheimer disease; flexibility; gamma secretase; member; neuron development; neuron loss; neuropsychiatric disorder; neurotransmitter release; particle; postsynaptic; presynaptic; prevent; programs; protein complex; receptor; reconstitution; screening; secretase; structural biology; synaptic function; synaptogenesis; thermostability

Project Summary/Abstract Alzheimer disease (AD) is a neurodegenerative disease characterized by progressive decline of brain functions, progressive loss of synapses, neuronal death, brain atrophy and formation of extracellular amyloid plaques and intracellular neurofibrillary tangles. The amyloid precursor protein (APP) is a type I transmembrane glycoprotein which plays a central role in AD pathogenesis, as the precursor of the beta amyloid peptide (Aβ), the main component of amyloid plaques. In addition, several mutations on the APP gene are associated with the early-onset hereditary form of the disease (Familial Alzheimer disease or FAD). Apart from being the precursor of the toxic Aβ peptide and the carrier of FAD mutations, APP has been associated with a wide range of biological functions, including proliferation, neuronal development, intracellular transport, Ca2+ homeostasis, neurite outgrowth, shaping of dendritic complexity and, most importantly, synaptogenesis and synaptic function. Indeed, multiple reports link APP to the regulation of spine density, spine dynamics and synaptic plasticity. Reported synaptic roles of APP both in the presynaptic active zone and postsynaptic density include regulation of presynaptic nicotinic receptors, facilitation of neurotransmitter release, regulation of synaptic vesicle recycling and regulation of the cell surface expression and activity of NMDA receptors. Deregulation of one or more of the above functions is therefore likely to lead to synaptic dysfunction and neurodegeneration, also basic hallmarks of the AD. APP is part of a gene family, with two more mammalian members, the Amyloid Precursor Like Protein 1 (APLP1) and the Amyloid Precursor Like Protein 2 (APLP2). Both APLPs are highly homologous to APP, with similar molecular architecture and functions, especially related to synaptogenesis, spine dynamics and synaptic plasticity. The goal of this proposal is to develop suitable experimental resources for performing structural investigations of the physiological functions of APP and APLPs. To achieve this goal, we propose to establish an expression and purification pipeline for medium throughput screening of APP, APLP1 and APLP2 orthologs, investigate an alternative tagging procedure and compare extraction and reconstitution methods for a subset of orthologs. Finally, we propose to utilize single particle cryo-electron microscopy (cryoEM) to elucidate atomic-level details of the molecular architecture of APP family proteins. High-resolution structural information of a full- length APP family protein will enable us to better understand the structural basis of the physiological functions of APP and APP-like proteins and inform our understanding of how these functions are deregulated in AD.

项目摘要/摘要 阿尔茨海默氏病（AD）是一种神经退行性疾病，其特征是大脑的逐渐下降 功能，突触的进行性丧失，神经元死亡，脑萎缩和细胞外淀粉样蛋白的形成 斑块和细胞内神经纤维缠结。淀粉样蛋白前体蛋白（APP）是I型 跨膜糖蛋白在AD发病机理中起着核心作用，作为β的前体 淀粉样蛋白肽（Aβ），淀粉样斑块的主要成分。此外，该应用程序上有几个突变 基因与该疾病的早期遗传形式（家族性阿尔茨海默氏病或​​FAD）有关。 除了是有毒Aβ辣椒的前体和FAD突变的载体外，App一直是 与广泛的生物学功能相关，包括增殖，神经元发展， 细胞内转运，Ca2+体内稳态，神经蛋白神经落生，树突复杂性的形状，大多数 重要的是，突触发生和突触功能。确实，多个报告链接应用程序与脊柱的调节 密度，脊柱动力学和合成可塑性。报道了应用程序在突触前活动中的合成作用 区域和突触后密度包括调节突触前烟碱受体，设施 神经递质释放，突触囊泡回收的调节和细胞表面表达的调节 NMDA接收器的活动。因此，放松一个或多个上述功能的管制可能会导致 为了突触功能障碍和神经变性，也是AD的基本标志。应用是基因家族的一部分， 还有两个哺乳动物成员，淀粉样蛋白前体（例如蛋白1（APLP1））和淀粉样蛋白 像蛋白2（APLP2）这样的前体。两个APLP都与App高度同源，具有相似的分子 建筑和功能，尤其与突触发生，脊柱动力学和突触可塑性有关。这 该建议的目标是开发合适的实验资源，以进行结构研究 应用程序和APLP的物理功能。为了实现这一目标，我们建议建立一个表达 以及用于APP，APLP1和APLP2直系同源物的中等吞吐量筛选的纯化管道，研究 替代标记程序，并比较直系同源物的一部分的提取和重新建立方法。 最后，我们建议利用单个粒子冷冻电子显微镜（冷冻）阐明原子级 App家族蛋白的分子结构的细节。完整的高分辨率结构信息 长度应用家庭蛋白将使我们能够更好地了解生理功能的结构基础 应用程序和类似应用的蛋白质，并告知我们对这些功能如何在AD中放松管制的理解。