Lipoprotein Quantitation and Function Core

摘要(核心C) 高质量、一致和可重现的量化和功能数据是这项研究成功的关键 计划项目。高密度脂蛋白量化和功能核心(核心C)的总体目标就是满足这一点 在整个计划中以集中且一致的方式满足客户的需求。核心的结构旨在提供 以下服务：1)高密度脂蛋白颗粒大小和浓度的定量(无论是从血浆还是从血浆中分离出来的 重构成的高密度脂蛋白颗粒)，用于特定结构研究，使用校准的差示离子迁移率分析 (CIMA)；2)质谱分析，以表征分离或分离的高密度脂蛋白颗粒的蛋白质组成 由项目调查人员准备，以及支持其结构研究的化学交联策略； CORE还将使用质谱学来确认由 核心D；3)通过校准IMA精确测量高密度脂蛋白颗粒浓度和 LC-MS/MS准确定量载脂蛋白，包括定量载脂蛋白化学计量 高密度脂蛋白颗粒上的载脂蛋白；4)用于解释高密度脂蛋白和蛋白质组数据集的生物信息学支持以及 用于将数据与来自基因本体论、蛋白质-蛋白质相互作用和途径的信息相结合 分析；5)使用幼年仓鼠肾细胞测量胆固醇和磷脂外流 有条件表达WT和突变人ABCA1。核心C将由Tomas Vaisar博士领导，正在工作 与唐崇仁博士密切合作，并将设在华盛顿大学南湖联盟 校园，也是项目1所在的地方。这两个实验室在所有领域都拥有广泛的、有据可查的专业知识 核心的建议服务。在之前的PPG周期中，该核心执行了1000多次高密度脂蛋白-P分析 和大小，500多个高密度脂蛋白蛋白质组的LC-MS/MS分析，以及用于表征和 实验验证了高密度脂蛋白和ABCA1结构的理论模型。这些服务将在 这项提议。总而言之，它们将使成本效益和严格的测试产生的假设 各种项目。

ABSTRACT (Core C) High quality, consistent and reproducible quantitation and functional data are critical for the success of this Program Project. The overarching goal of the HDL Quantitation and Function Core (Core C) is to meet this need in a centralized and consistent manner across the entire Program. The Core is structured to provide the following services: 1) quantitation of HDL particle size and concentration (whether isolated from plasma or reconstituted HDL particles) for specific structural studies, using calibrated differential ion mobility analysis (cIMA); 2) mass spectrometric analyses to characterize the protein composition of HDL particles isolated or prepared by Program investigators, and chemical crosslinking strategies to support their structural studies; the Core will also use mass spectrometry to confirm the identity and purity of the recombinant proteins prepared by Core D; 3) a combination of accurate measurement of HDL particle concentration by calibrated IMA and accurate quantification of apolipoproteins by LC-MS/MS, including quantification of the stoichiometry of apolipoproteins on HDL particles; 4) bioinformatic support for interpreting HDL-P and proteomic data sets and for combining the data with information from Gene Ontology, protein-protein interactions, and pathway analysis; 5) measurements of cholesterol and phospholipid efflux, using baby hamster kidney cells conditionally expressing WT and mutant human ABCA1. Core C will be led by Tomas Vaisar, Ph.D., working closely with Chongren Tang, Ph.D., and it will be located at the University of Washington South Lake Union campus, where Project 1 is also located. Both labs have extensive, well-documented expertise in all areas of the Core’s proposed services. In the previous PPG cycle, this Core performed over 1000 analyses of HDL-P and size, over 500 LC-MS/MS analyses of HDL’s proteome, and crosslinking studies for characterizing and experimentally verifying theoretical models of HDL and ABCA1 structures. These services will be extended in this proposal. Collectively, they will enable cost-effective and rigorous testing of the hypotheses generated by the various Projects.