Studies of p53 and Amyloidogenic Proteins

p53 和淀粉样蛋白的研究

• 批准号: 7255718
• 负责人: FEIMENG ZHOU
• 金额: $ 19.12万
• 依托单位: CALIFORNIA STATE UNIVERSITY LOS ANGELES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7255718
• 关键词: DNA; amyloid proteins; amyloidosis; atomic force microscopy; chemical aggregate; chemical kinetics; electrical measurement; electrical potential; electrochemistry; intermolecular interaction; ions; metalloproteins; metals; molecular assembly /self assembly; oxidation reduction reaction; p53 gene /protein; protein structure function; scanning tunneling microscopy; surface plasmon resonance; surface property

The central theme of this proposal is the redox-modulated protein interactions at the surface/solution interface. One subproject is concerned with the electrochemical studies of p53 (a tumor suppression protein) and the correlation of the externally applied potential to the extent of p53/DNA binding. The other subproject will deal with the redox reactions inherent in amyloidogenic proteins and their relationships with the protein fibrillation/aggregation processes. The proposed work will utilize a variety of analytical and surface techniques to study DNA/protein and protein/small molecule interactions. These techniques include, but are not limited to, voltammetry, flow injection quartz crystal microbalance, surface plasmon resonance, scanning tunneling microscopy, scanning electrochemical microscopy, and atomic force microscopy. Particular emphasis will be placed on the voltammetric and surface characterization of p53 protein and alpha-synuclein and beta-amyloid. Various immobilization schemes will be explored to immobilize proteins with optimal surface configuration (i.e., surface coverage and molecular orientation) and without significant conformational changes. Kinetic measurements will also be performed on the interaction of these species with their biological partners (e.g., binding of p53 to its consensus DNA under various applied potentials and interaction of potential therapeutic drugs with amyloidogenic proteins). The influence of various species (e.g., redox-active metal ions and organic molecules and electroactive metalloproteins) on the voltammetric behavior and binding kinetics of p53 and amyloidogenic proteins will also be investigated. The redox properties of these proteins will be correlated with findings from kinetic studies and surface characterization. Attempts will also be made in the identification of intermediates of amyloidogenic protein fibrillation and aggregation. The studies proposed herein should help elucidate the mechanisms involved in the interactions of the two important types of proteins with other species under physiologically relevant conditions and provide useful information to the effective development of therapeutic drugs for cancer and neurodegenerative diseases.

该提案的中心主题是表面/溶液界面处氧化还原调节的蛋白质相互作用。其中一个子项目涉及 p53（一种肿瘤抑制蛋白）的电化学研究以及外部施加电位与 p53/DNA 结合程度的相关性。另一个子项目将处理淀粉样蛋白固有的氧化还原反应及其与蛋白质原纤维化/聚集过程的关系。拟议的工作将利用各种分析和表面技术来研究 DNA/蛋白质和蛋白质/小分子相互作用。这些技术包括但不限于伏安法、流动注射石英晶体微天平、表面等离子体共振、扫描隧道显微镜、扫描电化学显微镜和原子力显微镜。将特别强调 p53 蛋白、α-突触核蛋白和 β-淀粉样蛋白的伏安法和表面表征。将探索各种固定方案以固定具有最佳表面构型（即表面覆盖和分子取向）并且没有显着构象变化的蛋白质。还将对这些物种与其生物伙伴的相互作用进行动力学测量（例如，在各种应用电位下 p53 与其共有 DNA 的结合以及潜在治疗药物与淀粉样蛋白的相互作用）。还将研究各种物种（例如氧化还原活性金属离子和有机分子以及电活性金属蛋白）对 p53 和淀粉样蛋白的伏安行为和结合动力学的影响。这些蛋白质的氧化还原特性将与动力学研究和表面表征的结果相关。还将尝试鉴定淀粉样蛋白原纤维化和聚集的中间体。本文提出的研究应有助于阐明两种重要类型的蛋白质在生理相关条件下与其他物种相互作用的机制，并为有效开发癌症和神经退行性疾病的治疗药物提供有用的信息。