Core C: GAGomics and Proteomics

核心 C：GAG 组学和蛋白质组学

• 批准号: 10088967
• 负责人: Adam M Hawkridge
• 金额: $ 25.93万
• 依托单位: VERSITI WISCONSIN, INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-01 至 2025-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10088967
• 关键词: Address; Affinity Chromatography; Area; Aspirate substance; Binding; Biological; Biological Process; Biology; Bone Marrow; Bone Marrow Cells; Capillary Electrophoresis; Carbohydrates; Cells; Chondroitin Sulfates; Clinical; Communities; Complement; Complex; Computer software; Coupled; Data; Data Analyses; Data Set; Deductibles; Disease; Environment; Extracellular Protein; Foundations; Genetic; Genetic Transcription; Genome; Glycobiology; Glycosaminoglycans; Glypican; Goals; Hematopoietic System; Hematopoietic stem cells; Heparin; Heparitin Sulfate; Label; Laboratories; Link; Liquid Chromatography; Mass Spectrum Analysis; Measures; Megakaryocytes; Membrane Proteins; Modification; Molecular; Mus; Names; Patients; Performance; Polysaccharides; Preparation; Protein Analysis; Proteins; Proteomics; Publishing; RNA; RPL29 gene; Research; Sampling; Serine; Stable Isotope Labeling; Structural Models; Structure; System; Techniques; Technology; Universities; Virginia; X ray spectroscopy; base; biophysical properties; carbohydrate structure; cell growth regulation; chondroitin sulfate glycosaminoglycan; comparative; data repository; extracellular; glycoproteomics; innovation; insight; instrumentation; perlecan; polysulfated glycosaminoglycan; programs; protein expression; secretory protein; serglycin; stem cell biology; syndecan; tool

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.

核心C -项目总结/摘要 糖胺聚糖（GAG）是一种线性O-连接的多糖，可对丝氨酸残基进行选择性修饰 膜蛋白（例如，磷脂酰肌醇蛋白聚糖：GPC 1-GPC 6，多配体聚糖：SDC 1-SDC 4），分泌蛋白（sergylycin）， 和细胞外蛋白质（例如，perlecan）。GAG以非模板导向的方式合成， 大的异质结构，不能以类似的方式预测，我们可以使用基因组， 预测转录（RNA）和翻译（蛋白质）结构/序列。因此，这些结构 很难从结构上描述。此外，将生物功能赋予这些结构 例如蛋白质-GAG相互作用增加了糖分析的挑战。高效质谱 LC-MS/MS结合先进的样品制备策略是结构分析的领先技术。 GAG结构的表征和定量。蛋白糖组学核心C（PG-核心C）填补了一个关键的 技术上的需要，已概述了三个具体目标的计划项目。具体目标 1、PG-Core C将对乙酰肝素和硫酸软骨素进行比较结构分析 来源于骨髓龛的糖胺聚糖。这个目标，我们称之为GAGomics， 为所有三种GAG修饰、细胞调节和蛋白质-GAG相互作用提供了一个全面的衡量标准。 项目在SPECIFIC AIM 2中，PG-Core C将对蛋白质-GAG进行深入的蛋白质组学分析 我们称之为GAGactomics的骨髓小生境中的相互作用， 差异结合细胞或细胞外GAG结构。在具体目标3中，或目标仅集中于 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。 在骨髓龛中表达。我们的总体目标是支持所有三个创新的假设 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， 蛋白质表达水平在骨髓龛。