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Imaging, Protein Production, and Chemical Biology Core

成像、蛋白质生产和化学生物学核心

基本信息

批准号：
10613966
负责人：
Loren Gi Fong
金额：
$ 51.48万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-05-01 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10613966
关键词：
Alkynes Antibodies Antibody Formation Azides Binding Binding Sites Biology Biomedical Research Biotin Cells Chemicals Chemistry Cholesterol Cholesterol Homeostasis Diazomethane Disease Electron Microscope Electron Microscopy Electrons Ensure Enzyme-Linked Immunosorbent Assay Equipment Fab Immunoglobulins Fluorescence Microscopy Funding Goals Health High Density Lipoproteins Human Resources Image Insecta Investigation Isotopes Label Light Lipids Mass Spectrum Analysis Microscopy Movement National Heart, Lung, and Blood Institute Preparation Production Productivity Proteins Proteolysis Proteome Proteomics Protocols documentation Reagent Recombinant Antibody Recombinant Proteins Regulation Research Research Personnel Sampling Scanning Electron Microscopy Services Signal Transduction Stable Isotope Labeling Sterols Structure System Tertiary Protein Structure Tissues Transmission Electron Microscopy Triglyceride Metabolism Visualization activity-based protein profiling cellular imaging cost crosslink cycloaddition equipment acquisition experience high resolution imaging imaging facilities imaging modality instrument light microscopy lipid transport mass spectrometer microscopic imaging mutant nanomachine novel novel strategies operation protein expression protein purification recruit sample collection sample fixation tool trafficking

项目摘要

Core A: Imaging, Protein Production, and Chemical Biology Core SUMMARY/ABSTRACT The goal of the Imaging, Protein Production, and Chemical Biology Core is to maximize the scientific progress and productivity of PPG investigators by providing essential services and reagents. The core will perform three major functions for the PPG: (1) advanced light and electron microscopy imaging;; (2) protein and antibody production;; and (3) chemical proteomics. All three PPG component projects will utilize at least two core services, and two of the projects will utilize all three. Centralizing these activities within a core will benefit the PPG by increasing productivity and reducing costs. The core will provide full-service scanning and transmission electron microscopy, standard and live-cell confocal fluorescence microscopy, and NanoSIMS imaging. The latter is a mass spectrometry–based imaging modality that makes it possible to create high-resolution images of cells and tissues based solely on their isotopic content. PPG investigators will use NanoSIMS to visualize and quantify the localization and movement of lipids in cells and tissues. PPG investigators will also study new molecules and novel mechanisms regulating lipid transport and signaling, for which few reagents exist. To assist these investigations, the core will produce protein and antibody reagents for PPG studies (e.g., soluble GPIHBP1, stable isotope–labeled ALO-D4, 5D2 Fab fragments). Finally, the core will perform Activity-Based Chemical Proteomics and isotopic Tandem Orthogonal Proteolysis‒Activity-Based Protein Profiling. These state-of-the-art proteomics discovery platforms will be used to identify proteins involved in the intracellular trafficking of HDL- derived cholesterol and the regulation of STING signaling by cholesterol. For the next five years, the core is poised—with the experience, robust protocols, and experienced personnel—to provide vital services to PPG investigators.

核心A：成像，蛋白质生产和化学生物学核心总结/摘要成像，蛋白质生产和化学生物学核心的目标是最大限度地提高科学进步通过提供基本服务和试剂，提高PPG调查人员的生产力。核心将执行三项任务 PPG的主要功能：（1）先进的光学和电子显微镜成像;（2）蛋白质和抗体 PPG的所有三个组成项目将利用至少两个核心服务，将这些活动集中在一个核心内将有利于项目编制补助金，提高生产率，降低成本。核心将提供全方位服务的扫描和透射电子显微镜，标准和活细胞共聚焦荧光显微镜，和NanoSIMS成像。后者是一个 - 基于质谱的成像模式，其使得可以创建细胞的高分辨率图像， PPG研究人员将使用NanoSIMS来可视化和量化组织中的同位素含量。细胞和组织中脂质的定位和运动。PPG研究人员还将研究新的分子，调节脂质转运和信号传导的新机制，其试剂很少。研究，核心将生产用于PPG研究的蛋白质和抗体试剂（例如，可溶性GPIHBP 1，最后，核心将进行基于活性的化学分析，蛋白质组学和同位素串联正交蛋白质水解酶活性分析的基础上的蛋白质谱。这些国家的最先进的蛋白质组学发现平台将用于鉴定参与细胞内高密度脂蛋白转运的蛋白质衍生胆固醇和胆固醇调节STING信号。在未来五年，核心是具备丰富的经验、强大的协议和经验丰富的人员，为PPG提供重要服务 investigators.