Physical Biochemistry and Biology of Amyloid Beta-Protein

β-淀粉样蛋白的物理生物化学和生物学

• 批准号: 8722423
• 负责人: DAVID B. TEPLOW
• 金额: $ 31.57万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-15 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8722423
• 关键词: Alzheimer' s Disease; Amyloid; Amyloid Proteins; Amyloid beta-Protein; Biochemistry; Bioinformatics; Biological; Biology; Cell Line; Cereals; Circular Dichroism; Country; Development; Disease; Drosophila eye; Eye Development; Fluorescence; Foundations; Genes; Goals; Human; Hydroxyl Radical; In Vitro; Lead; Life; Locomotion; Longevity; Mass Spectrum Analysis; Modeling; Molecular Conformation; Mutagenesis; Neurons; Neurotoxins; Pathologic; Peptides; Printing; Proteins; Proteolysis; Research; Resolution; Scanning; Seeds; Seminal; Solvents; Spectrum Analysis; Structure; Structure-Activity Relationship; System; Therapeutic; Therapeutic Agents; Thioflavin T; Time; Tissue-Specific Gene Expression; United States; Work; aged; crosslink; cytotoxic; foot; in vivo; insight; ion mobility; light scattering; molecular dynamics; multidisciplinary; neurotoxic; novel; peptide A; prevent; research clinical testing; senescence; structural biology; tau Proteins

DESCRIPTION (provided by applicant): Alzheimer's disease (AD) is a devastating disease of the aged. Two amyloid-forming proteins are associated with AD, the amyloid ?-protein (A?) and tau. Recent evidence supports an hypothesis, the "amyloid cascade hypothesis," that posits that A? oligomers are the seminal neuropathogenetic agents in AD. The overall goal of this proposal is to understand the structural biology of A?, and fragments thereof, and to establish formal structure-activity relationships. In the long run, we seek to obtain an atomic-resolution determination of the structure of the proximate neurotoxins formed by A?, and in doing so, enable the development, for the first time, of disease-modifying AD treatments. A multidisciplinary strategy, employing complementary experimental and computational approaches, will be employed. This strategy has been used very successfully in the past, providing novel insights into the A? system. Three specific aims are proposed that systematically and logically progress from in vitro biophysical studies of A? and its oligomeric assemblies (Aim 1), to in vitro and in vivo studies of the biological activity of selected such assemblies (Aim 2), to determination of the effects of selected assemblies on differential gene expression in neurons (Aim 3). Taken together, these studies will provide the theoretical and experimental foundation for subsequent therapeutic compound development and clinical testing in humans. Aim 1. To determine the structural dynamics of A? and tau assembly. a. To use scanning Tyr mutagenesis to elucidate mechanisms of A? oligomerization. b. To determine the dynamics of intramolecular turn formation and its effects on A? assembly. c. To determine the effects of primary structure changes on the conformations and assembly dynamics of biologically relevant and theoretically important A? peptides. Aim 2. To determine the biological effects of A? assemblies. a. To determine the cytotoxic effects of A? assemblies on cultured neuronal cell lines and primary neurons. b. To determine the effects of A? assemblies on Drosophila eye development, locomotion, and longevity. Aim 3. To identify and validate AD-relevant genes using a bioinformatics approach that considers A? assembly structure, neuron type, and neuron senescence.

描述（由申请人提供）：阿尔茨海默病（AD）是一种毁灭性的老年人疾病。两种淀粉样蛋白形成蛋白与 AD 相关，即淀粉样蛋白 β-蛋白 (A?) 和 tau。最近的证据支持一个假设，即“淀粉样蛋白级联假说”，该假说假设 A？寡聚体是 AD 中的重要神经致病因子。该提案的总体目标是了解Aβ及其片段的结构生物学，并建立正式的结构-活性关系。从长远来看，我们寻求获得Aβ形成的近端神经毒素结构的原子分辨率测定，并在此过程中首次实现改善疾病的AD治疗方法的开发。将采用多学科策略，采用互补的实验和计算方法。这一策略在过去已经被非常成功地使用，为 A? 提供了新颖的见解。系统。提出了三个具体目标，系统地、逻辑地从 A? 的体外生物物理研究中取得进展。及其寡聚组装体（目标 1），对选定此类组装体的生物活性进行体外和体内研究（目标 2），确定选定组装体对神经元差异基因表达的影响（目标 3）。总而言之，这些研究将为后续治疗化合物的开发和人体临床测试提供理论和实验基础。目标 1. 确定 A? 的结构动力学？和 tau 组装。一个。使用扫描 Tyr 诱变来阐明 A？寡聚化。 b.确定分子内转角形成的动力学及其对 A? 的影响？集会。 c.确定一级结构变化对生物学相关且理论上重要的 A 的构象和组装动力学的影响？肽。目标 2. 确定 A? 的生物学效应组件。一个。确定A的细胞毒性作用？培养的神经元细胞系和原代神经元上的组装。 b.确定 A 的效果？果蝇眼睛发育、运动和寿命的组装。目标 3. 使用考虑 A? 的生物信息学方法来识别和验证 AD 相关基因。装配结构、神经元类型和神经元衰老。