STRUCTURAL AND FUNCTIONAL PROPERTIES

结构和功能特性

基本信息

批准号：
7877908
负责人：
JOYCE LUSTBADER
金额：
$ 31.79万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2000
资助国家：
美国
起止时间：
2000-09-01 至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/7877908
关键词：
Active Sites Aging Alcohols Alzheimer&apos s Disease Amyloid Attenuated Binding Binding Sites Biology Cells Cellular Stress Chemosensitization Complex Computer Assisted Crystallography Cyclophilins Data Data Analyses Environment Enzymes Free Radical Induced Apoptosis Pathway Free Radicals Functional disorder Future Generations Goals Grant Immunoprecipitation In Vitro Isoelectric Focusing Laboratories Link Maps Mediating Memory Methodology Mitochondria Mitochondrial Matrix Modeling Mus Neurons Nicotinamide adenine dinucleotide Oxidative Stress Oxidoreductase Pathogenesis Pathology Pathway interactions Patients Peptide Fragments Peptides Plasma Point Mutation Property Reagent Research Personnel Role Site Stress Structure Surface Synapses Techniques Testing Therapeutic Toxic effect Transgenic Mice Transgenic Organisms Work aqueous base cofactor crosslink cyclophilin D cytotoxicity in vivo inhibitor/antagonist insight mitochondrial dysfunction mitochondrial permeability transition pore mutant novel therapeutic intervention overexpression peptide A peptide B pharmacophore polypeptide prevent programs yeast two hybrid system

项目摘要

Amyloid-B peptide (AB) is believed to have a central role in the pathogenesis of Alzheimer's disease (AD). Intracellular mechanisms by which AB disrupts cellular properties, leading to neuronal and synaptic dysfunction, and mitochondrial damage, are not well understood. ABAD (A6 Binding Alcohol Dehydrogenase) is an intracellular enzyme with unique features including a wide range of substrates, binding AB in the nanomolar range, and exacerbating AB-induced cell stress. Recent results from this laboratory and others support a connection between ABAD and AB-mediated mitochondrial and synaptic dysfunction (project 3) relevant to AD pathology. Ap interacts specifically with ABAD in the mitochondria of AD patients and transgenic mice. Transgenic mice overexpressing ABAD in an Ap-rich milieu present exaggerated neuronal oxidative stress, mitochondrial dysfunction, and impaired memory. The crystal structure of Ap-bound ABAD shows substantial deformation of the active site that prevents nicotinamide adenine dinucleotide (NAD) binding. From the crystal structure, the putative AB interaction site in ABAD spans residues 94-114. A peptide encompassing this region of ABAD, residues 91-119, specifically inhibits the ABAD-Ap interaction and suppresses Ap-induced apoptosis and free- radical generation in neurons. Indicating that binding of ABAD to AB is crucial in AB-mediated neuronal stress. Possible mechanisms underlying ABAD-mediated changes in mitochondrial properties are indicated by our findings that AB inhibits ABAD enzymatic activity, the observation that ABAD binds cyclophilin (cypD), sequestering the latter in the mitochondrial matrix, and potential toxic effects of ABAD-ABcomplex (e.g., generation of ROS). This competitive renewal is founded on the hypothesis that ABAD is a critical intracellular target potentiating AB-mediated mitochondrial dysfunction and, thus, leading to cellular perturbation in an AB-rich environment. Blockade of the ABAD-ABinteraction may be a potential therapeutic approach in the treatment of AD. The aims of Project 4 are to identify the basis of ABAD-AB, ABAD-cyclophilin D (cypD) interactions using crystal structure techniques, and to analyze critical aspects of the intracellular pathway through which ABAD engages AB, cypD, and ABAD-AB complex induces cellular stress. Project 4 will work closely with Projects 1-3, and will obtain technical assistancefrom CoreA. Collaborative interactions will include: sharing of reagents for analysis of ABAD (Project 3), mechanisms for evaluating cell stress (Projects 1-3), and assistance in data analysis (core A).

B 淀粉样蛋白肽 (AB) 被认为在阿尔茨海默病的发病机制中发挥着核心作用（广告）。 AB 破坏细胞特性的细胞内机制，导致神经元和突触功能障碍和线粒体损伤尚不清楚。 ABAD（A6 结合醇脱氢酶）是一种具有独特功能的细胞内酶，包括多种底物，在纳摩尔范围内结合 AB，并加剧 AB 诱导的细胞应激。最近的结果实验室和其他人支持 ABAD 和 AB 介导的线粒体和与 AD 病理学相关的突触功能障碍（项目 3）。 Ap 专门与 ABAD 相互作用 AD患者和转基因小鼠的线粒体。富含 Ap 的转基因小鼠过度表达 ABAD 环境呈现过度的神经元氧化应激、线粒体功能障碍和记忆力受损。 Ap 结合的 ABAD 的晶体结构显示出活性位点的显着变形，从而阻止了烟酰胺腺嘌呤二核苷酸 (NAD) 结合。从晶体结构来看，AB ABAD 中的相互作用位点跨越残基 94-114。包含 ABAD 这个区域的肽，残基 91-119，特异性抑制 ABAD-Ap 相互作用并抑制 Ap 诱导的细胞凋亡和游离神经元中自由基的产生。表明 ABAD 与 AB 的结合在 AB 介导的过程中至关重要神经元应激。 ABAD 介导的线粒体特性变化的可能机制我们的发现表明 AB 抑制 ABAD 酶活性，观察到 ABAD 结合亲环蛋白 (cypD)，将后者隔离在线粒体基质中，并产生潜在的毒性作用 ABAD-AB复合体（例如，ROS的产生）。这种竞争性更新是建立在以下假设之上的： ABAD 是一个关键的细胞内靶标，可增强 AB 介导的线粒体功能障碍，并且因此，在富含 AB 的环境中导致细胞扰动。封锁 ABAD-AB 相互作用可能是治疗AD的潜在治疗方法。项目 4 的目标是确定使用晶体结构技术建立 ABAD-AB、ABAD-亲环蛋白 D (cypD) 相互作用的基础，并分析 ABAD 参与 AB、cypD 和的细胞内途径的关键方面 ABAD-AB 复合物诱导细胞应激。项目 4 将与项目 1-3 密切合作，并将从 CoreA 获得技术援助。协作互动将包括：共享试剂 ABAD 分析（项目 3）、评估细胞应激的机制（项目 1-3）以及协助数据分析（核心A）。