Nascent Dynamic Intermediates in Amyloid Aggregation

淀粉样蛋白聚集中的新生动态中间体

基本信息

批准号：
6963283
负责人：
WATT W WEBB
金额：
$ 20.15万
依托单位：
CORNELL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-08-15 至 2007-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6963283
关键词：
alpha synuclein amyloid proteins conformation cytotoxicity electrical property fluorescence microscopy fluorescence polarization fluorescence spectrometry intermolecular interaction lipid bilayer membrane myoglobin phase change protein folding protein protein interaction protein structure function sedimentation equilibrium

项目摘要

DESCRIPTION (provided by applicant): The long-term goal of this proposal is development of a detailed kinetic description of the initiation of amyloid aggregation, a common feature of the diverse group of devastating diseases characterized by the deposition of fibrillar aggregates of misfolded proteins. The focus is a description of the identities and functions of the nascent intermediate molecular species as a means for identifying target protein species for the development of methods of controlling the aggregation pathway. The specific proposed hypothesis is that non-fibrillar oligomeric protein species in conjunction with conformational changes in the native proteins play the crucial role in the initiation of aggregation of amyloid-forming proteins. This hypothesis is based on evidence that 1) soluble oligomers of amyloid proteins are known to be cytotoxic and 2) solutions influencing protein secondary structure of amyloid proteins affect their aggregation kinetics. The experimental strategies rely on modern biophysical instrumentations based on multiphoton microscopy and fluorescence correlation spectroscopies to characterize the solution and lipid-associated behavior of three key amyloid-forming proteins: alpha-Synuclein, A-beta, and apomyoglobin. Two specific aims are: 1. Quantification and understanding of the conformational changes associated with the initiation of aggregation utilizing adaptations of fluorescence correlation spectroscopy to characterize protein conformational fluctuations and folding/misfolding associated conformational changes in apomyoglobin, alpha-Synuclein, and A-beta. 2. Biophysical characterization of the soluble oligomers formed by alpha- Synuclein and A-beta, including (i) their solution equilibrium properties, and (ii) the nature of the interactions with lipid membranes, in particular the role of membrane charge and lipid-ordered phase formation, to understand how cell membrane interactions affect aggregation.

描述（由申请人提供）：该提案的长期目标是开发淀粉样蛋白聚集的启动的详细动力学描述，淀粉样蛋白的启动，这是多种毁灭性疾病的共同特征，其特征是折叠蛋白异折叠蛋白的原纤维聚集体的沉积。重点是对新生中间分子物种的身份和功能的描述，作为识别靶蛋白质物种以开发控制聚集途径的方法的一种手段。提出的具体假设是，非纤维寡聚蛋白种类与天然蛋白的构象变化结合使用，在淀粉样蛋白蛋白的聚集中起着至关重要的作用。该假设基于证据：1）已知淀粉样蛋白的可溶性低聚物是细胞毒性的，而2）影响淀粉样蛋白的蛋白二级结构的溶液会影响其聚集动力学。实验策略依赖于基于多光子显微镜和荧光相关光谱的现代生物物理仪器来表征三种关键淀粉样蛋白蛋白的溶液和脂质相关的行为：α-核蛋白，a-beta，a-beta，a-beta和apomyoglobin。两个具体的目的是：1。使用荧光相关光谱适应的聚集构象变化的定量和理解，以表征蛋白质构象波动以及倍肌球蛋白，α-核素和a-β的蛋白质构象波动以及折叠/折叠/错误的构象变化。 2。由α-突触核蛋白和a-beta形成的可溶性低聚物的生物物理表征，包括（i）它们的溶液平衡特性，以及（ii）与脂质膜相互作用的性质，特别是膜电荷和脂质 - 脂质和脂质 - 有序的相形成，以了解细胞膜相互作用如何影响聚集。