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总结 该 P01 提案的总体目标是阐明蛋白质生产和化学生物学。 钙化机制及其在年龄相关性黄斑的发病和病因学中的潜在作用 拟议的项目将研究退化（AMD）和阿尔茨海默病（AD）。 分子、细胞和生物水平，以建立对责任的全面看法 对于这种与衰老相关的现象，所有这些努力的核心将是提供纯净且同质的物质。 具有明确特性和修饰的蛋白质试剂，可以在各种测定中进行操作 该核心项目的目标是设计和制备这些试剂。 特别关注钙化相关蛋白玻连蛋白 (Vn)、Tau 和 β 淀粉样蛋白 (Aβ)。 还将致力于开发用于生产结构和成像修饰蛋白的增强工具 更具体地说，核心将致力于实现以下目标：1）Vn、Tau 和 Aβ 的生产。 构建所有项目所需的结构、生化、生物物理和细胞分析 提案2）开发基于内含肽的核磁蛋白质分段同位素标记工具 项目 1 中所述的共振 (NMR) 研究；以及 3) 荧光标记蛋白质的制备 除了为本文中描述的项目提供服务外，还为所有项目提供生物物理测定和传感器开发。 根据提案，我们期望核心开发的方案和试剂将成为化学研究的宝贵工具。 以及细胞生物学家、生物物理学家和结构生物学家，研究广泛的健康生物学问题 和疾病。