Core B - Biomolecular tools

核心 B - 生物分子工具

• 批准号: 10628927
• 负责人: Francesca M Marassi
• 金额: $ 37万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10628927
• 关键词: Age related macular degeneration; Aging; Alkaline Phosphatase; Alzheimer' s Disease; Amyloid beta-Protein; Biochemical; Biological; Biological Assay; Biology; Biophysics; Cells; Chemicals; Chemistry; Cysteine; Data Analyses; Development; Diphosphates; Disease; Drusen; Escherichia coli; Etiology; Event; Eye; Fluorescence Spectroscopy; Goals; Health; Hydroxyapatites; Image; In Vitro; Isotope Labeling; Label; Length; Lipids; Mammalian Cell; Methods; Modification; Molecular; Mus; Nuclear Magnetic Resonance; Nuclear Proteins; Participant; Patients; Phosphorylation; Post-Translational Protein Processing; Preparation; Process; Production; Property; Proteins; Protocols documentation; Reagent; Role; Sampling; Structural Biologist; Structure of retinal pigment epithelium; Testing; Time; Tissues; Validation; Vitronectin; Work; calcification; data structure; design; experimental study; imaging study; inorganic phosphate; intein; novel; particle; sensor; structural biology; tau Proteins; tool

CORE B SUMMARY Protein Production and Chemical Biology. The overall goal of this P01 proposal is to elucidate the biological mechanisms of calcification and their potential role in the onset and etiology of age-related macular degeneration (AMD) and Alzheimer’s disease (AD). The proposed projects will investigate calcification at the molecular, cellular and organismal levels to build a comprehensive view of the participants that are responsible for this aging-related phenomenon. Central to all these efforts will be the availability of pure and homogenous protein reagents with defined properties and modifications that can be manipulated in the various assays and experiments proposed in the projects. The goal of this Core will be to design and prepare these reagents, with a particular focus on the calcification-associated proteins vitronectin (Vn), Tau and amyloid β (Aβ). The Core will also aim to develop enhanced tools for the production of modified proteins for structural and imaging studies. More specifically, the Core will work towards the following goals: 1) Production of Vn, Tau, and Aβ constructs for structural, biochemical, biophysical and cellular analysis as necessary for all projects in the proposal; 2) Development of intein-based tools for segmental isotopic labeling of proteins for nuclear magnetic resonance (NMR) studies as described in Project 1; and 3) Preparation of fluorescently labeled proteins for biophysical assays and sensor development for all projects. In addition to serving the projects described in this proposal, we expect that the protocols and reagents developed by the Core will be valuable tools for chemical and cell biologists, biophysicists and structural biologists studying a wide range of biological problems in health and disease.

核心B摘要 蛋白质生产和化学生物学。该P01提案的总体目标是阐明生物学 计算机制及其在与年龄相关的黄斑的发作和病因中的潜在作用 变性（AMD）和阿尔茨海默氏病（AD）。拟议的项目将调查钙化 分子，细胞和有机水平，以建立负责参与者的全面观点 对于这种与衰老有关的现象。所有这些努力的中心将是纯和同质的可用性 具有定义特性和修饰的蛋白质试剂，可以在各种测定中操纵 项目中提出的实验。该核心的目的是设计和准备这些试剂，并 特别关注与计算相关蛋白玻璃体（VN），TAU和淀粉样蛋白β（Aβ）的关注。核心 还将旨在开发增强的工具，用于生产用于结构和成像的修饰蛋白 研究。更具体地说，核心将实现以下目标：1）生产VN，TAU和Aβ 必要的结构，生化，生物物理和细胞分析的构造 提议; 2）开发基于内部的基于内部的用于核磁蛋白的分段同位素标记的工具 共振（NMR）研究如项目1所述； 3）制备荧光标记的蛋白质 所有项目的生物物理测定法和传感器开发。除了服务于此所述的项目 提案，我们预计核心开发的协议和试剂将是化学的有价值工具 以及细胞生物学家，生物物理学家和结构生物学家，他们研究了各种各样的生物学问题 和疾病。