Structural characterization of APP family proteins

APP 家族蛋白的结构表征

基本信息

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
项目类别：
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&apos 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&apos 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

项目摘要

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.

项目概要/摘要阿尔茨海默病（AD）是一种神经退行性疾病，其特征是大脑进行性衰退功能、突触逐渐丧失、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原纤维缠结淀粉样前体蛋白 (APP) 是 I 型。跨膜糖蛋白，作为 β 的前体，在 AD 发病机制中发挥核心作用淀粉样肽（Aβ），淀粉样蛋白斑的主要成分此外，APP 上也有一些突变。基因与该疾病的早发遗传形式（家族性阿尔茨海默病或 FAD）有关。 APP除了是有毒Aβ肽的前体和FAD突变的载体之外，还被与广泛的生物功能相关，包括增殖、神经发育、细胞内运输、Ca2+稳态、神经突生长、树突复杂性的形成以及大多数重要的是，突触发生和突触功能确实有多个报告将 APP 与脊柱的调节联系起来。 APP 在突触前活动中的突触作用。区域和突触后密度包括突触前烟碱受体的调节，促进神经递质释放、突触小泡回收的调节和细胞表面表达的调节因此，一种或多种上述功能的失调可能会导致 NMDA 受体的活性。突触功能障碍和神经变性，也是 AD 的基本特征，也是基因家族的一部分。还有两个哺乳动物成员：淀粉样蛋白前体样蛋白 1 (APLP1) 和淀粉样蛋白前体样蛋白 2 (APLP2) 与 APP 高度同源，具有相似的分子。结构和功能，特别是与突触发生、脊柱动力学和突触可塑性相关。该提案的目标是开发合适的实验资源来进行结构研究 APP 和 APLP 的生理功能为了实现这一目标，我们建议建立一个表达。用于中通量筛选 APP、APLP1 和 APLP2 直向同源物的纯化管道，研究替代标记程序并比较直系同源子集的提取和重构方法。最后，我们建议利用单粒子冷冻电子显微镜（cryoEM）来阐明原子水平 APP 家族蛋白分子结构的详细信息。 APP家族蛋白的长度将使我们更好地了解其生理功能的结构基础 APP 和 APP 样蛋白的研究，并让我们了解这些功能在 AD 中是如何失调的。