Core C: GAGomics and Proteomics

核心C--项目摘要/摘要 糖胺多聚糖(GAG)是一种线性的O-连接的多糖，可以修饰特定数量的丝氨酸残基 膜蛋白(例如，Glypicans：GPC1-GPC6，Syndecans：SDC1-SDC4)，分泌蛋白(丝氨酸)， 和胞外蛋白(例如，Perlecan)。以非模板导向的方式合成GAG 无法用类似的方式预测的大型异质结构，我们可以使用基因组来 预测转录(RNA)和翻译(蛋白质)结构/序列。因此，这些结构 在结构上是非常难描述的。此外，赋予这些结构生物学功能 例如蛋白质-Gag的相互作用增加了糖分析的挑战。高性能质谱仪 (LC-MS/MS)结合先进的样品前处理技术是结构分析的领先技术 GAG结构的表征和量化。蛋白质组学-核心C(PG-核心C)填补了关键的 本计划项目在技术上的需要已在三个具体目标中概述。在特定目标中 1、PG-Core C将对肝素和硫酸软骨素进行比较结构分析 源于骨髓龛的糖胺多聚糖。这一目标，我们称之为GAG组学，将 为这三种生物提供GAG修饰、细胞调节和蛋白质-GAG相互作用的全球测量 项目。在特定的AIM 2中，PG-Core C将对蛋白质-Gag进行深入的蛋白质组学分析 在骨髓龛中的相互作用，我们称之为GAGacsecics或鉴定蛋白质 不同地结合细胞内或细胞外的GAG结构。在具体的目标3中，或目标仅侧重于 蛋白质组学，PG-Core C将对蛋白质进行全球和有针对性的定量蛋白质组学分析 在骨髓细胞中的表达。我们的总体目标是支持这三个方面的创新假设 具有最高质量LC-MS/MS数据的项目，以阐明GAG结构、GAG-蛋白质相互作用以及 骨髓穴中的蛋白质表达水平。

Core C – Project Summary/Abstract Glycosaminoglycans (GAGs) are linear O-linked polysaccharides that modify serine residues for a select number of membrane proteins (e.g., glypicans: GPC1-GPC6, syndecans: SDC1-SDC4), secretory proteins (serglycin), and extracellular proteins (e.g., perlecan). GAGs are synthesized in a non-template directed fashion producing large heterogeneous structures that cannot be predicted in an analogous way that we can use the genome to predict transcriptional (RNA) and translational (proteins) structures/sequences. Consequently these structures are exceedingly difficult to structurally characterize. Furthermore, assigning biological function to these structures such as protein-GAG interactions adds to the glycoanalytical challenges. High-performance mass spectrometry (LC-MS/MS) combined with advanced sample preparation strategies is the leading technology for structural characterization and quantification of GAG structures. The Proteoglycomics-Core C (PG-Core C) fills a critical technologically need for the Program Project which have been outlined in three Specific Aims. In SPECIFIC AIM 1, the PG-Core C will perform comparative structural analysis of heparan and chondroitin sulfate glycosaminoglycans derived from the bone marrow niche. This aim, which we have termed GAGomics, will provide a global measure of GAG modifications, cellular regulation, and protein-GAG interactions for all three Projects. In SPECIFIC AIM 2, the PG-Core C will perform in-depth proteomics analysis of protein-GAG interactions in the bone marrow niche which we've termed GAGactomics or the identification of proteins that differentially bind cellular or extracellular GAG structures. In SPECIFIC AIM 3, or the aim focused solely on Proteomics, the PG-Core C will perform global and targeted quantitative proteomics analysis of protein expression in the bone marrow niche. Our overall goal is to support the innovative hypotheses of all three Projects with the highest quality LC-MS/MS data to elucidate GAG structures, GAG-protein interactions, and protein expression levels in the bone marrow niche.