Core B - Biomolecular tools

核心B摘要 蛋白质生产和化学生物学。本P01提案的总体目标是阐明 钙化机制及其在老年性黄斑病变发病和病因学中的潜在作用 退行性变（AMD）和阿尔茨海默病（AD）。拟议的项目将调查钙化在 分子、细胞和有机体水平，以全面了解负责的参与者 这种与衰老有关的现象。所有这些努力的核心将是获得纯的和同质的 具有确定的性质和修饰的蛋白质试剂，其可以在各种测定中操作， 项目中提出的实验。本核心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和淀粉样蛋白β（Aβ）。核心 还将致力于开发用于生产结构和成像用改性蛋白质的增强工具 问题研究更具体地说，核心将朝着以下目标努力：1）生产Vn、Tau和Aβ 结构，生物化学，生物物理和细胞分析所需的所有项目的建设， 2）开发基于内含肽的工具，用于核磁共振的蛋白质片段同位素标记 如项目1中所述的核磁共振（NMR）研究;和3）制备荧光标记的蛋白质， 为所有项目提供生物物理分析和传感器开发。除了服务于本报告所述的项目外， 我们希望核心开发的方案和试剂将成为化学分析的宝贵工具。 细胞生物学家、生物药理学家和结构生物学家研究健康方面的各种生物学问题 和疾病

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.