Lipoprotein Quantitation and Function Core

摘要（核心C） 高质量、一致性和可重复的定量和功能数据是成功的关键。 计划项目。HDL定量和功能核心（核心C）的首要目标是满足这一点 在整个计划中以集中和一致的方式满足需求。核心的结构是提供 以下服务：1）HDL颗粒大小和浓度的定量（无论是从血浆中分离的还是从血液中分离的） 重组HDL颗粒）进行特定的结构研究，使用校准的微分离子迁移率分析 （cIMA）; 2）质谱分析以表征分离的HDL颗粒的蛋白质组成，或 由计划研究人员编写，并采用化学交联策略来支持其结构研究; 核心还将使用质谱法来确认通过以下方法制备的重组蛋白的身份和纯度： 核心D; 3）通过校准的IMA精确测量HDL颗粒浓度的组合， 通过LC-MS/MS准确定量载脂蛋白，包括定量载脂蛋白的化学计量 HDL颗粒上的载脂蛋白; 4）用于解释HDL-P和蛋白质组学数据集的生物信息学支持， 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）使用幼仓鼠肾细胞测量胆固醇和磷脂流出 条件性表达WT和突变型人ABCA 1。核心C将由Tomas Vaisar博士领导，工作 与汤崇仁博士密切合作，它将位于华盛顿南湖联盟大学 校园，项目1也位于。这两个实验室都拥有广泛的、有据可查的专业知识， 核心的服务。在上一个PPG周期中，该中心进行了1000多项HDL-P分析 和大小，超过500个HDL蛋白质组的LC-MS/MS分析，以及用于表征和 实验验证HDL和ABCA 1结构的理论模型。这些服务将于 这个提议。总的来说，它们将使人们能够以成本效益高的方式严格检验 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.