Rapid Sequencing of Sulfated Glycans By CE-MS/MS

通过 CE-MS/MS 对硫酸化聚糖进行快速测序

• 批准号: 9336350
• 负责人: I JONATHAN AMSTER
• 金额: $ 29.95万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9336350
• 关键词: Age; Anabolism; Animals; Binding Proteins; Biological; Biological Assay; Capillary Electrophoresis; Carbohydrates; Cations; Cessation of life; Chemistry; Communities; Complex; Coupling; DNA; Detection; Development; Disaccharides; Disease; Enzymatic Biochemistry; Enzymes; Event; Fluorescence; Glycosaminoglycans; Goals; Health; Heparin; Hexuronic Acids; High Pressure Liquid Chromatography; High-Throughput Nucleotide Sequencing; Human; Inflammation Process; Investigation; Ions; Length; Life; Light; Location; Lyase; Malignant Neoplasms; Mammalian Cell; Mass Spectrum Analysis; Measures; Methods; Modeling; Modification; Molecular; Monosaccharides; Nature; Oligosaccharides; Organism; Parents; Patients; Performance; Pharmacologic Substance; Physiology; Polysaccharides; Protein Binding Domain; Proteins; RNA; Recording of previous events; Records; Research; Resolution; Secretory Vesicles; Series; Speed; Structure; Structure-Activity Relationship; Surface; System; Techniques; Technology; Time; Tissues; Unspecified or Sulfate Ion Sulfates; Uronic Acids; base; chemical cleavage; insight; instrumentation; interest; mast cell; novel; polysulfated glycosaminoglycan; prototype; stereochemistry; success; sulfation; tandem mass spectrometry; tool; virtual

Project Summary/Abstract Sulfated glycosaminoglycan (GAG) carbohydrates represent one of the most structurally diverse groups of biomolecules and a comprehensive understanding of their biological structure-function relationships has yet to be achieved. Unlike other biomolecules such as DNA/RNA and proteins that are synthesized based upon a template, GAG biosynthesis is the result of the cumulative actions of a series of biosynthetic enzymes that produce a dynamic, polydisperse mixture. The composition of this mixture is dependent on factors such as organism age, developmental or disease state, and tissue of origin. Although this diversity presents a daunting analytical challenge, significant progress has been made in the field through attempts to isolate and characterize GAGs ranging from intact polysaccharides to enzymatically prepared oligosaccharides and disaccharides. The most widespread approach is to profile GAG disaccharides through separation (HPLC, UHPLC, HILIC, CE) and detection (UV, fluorescence, mass spectrometry (MS)). Domains can be characterized for structure-function studies by combining these techniques into hyphenated methods (e.g. LC- MS). Even though disaccharide analysis is the most widely utilized method for GAG analysis, it is unable to provide the structural motifs in GAGs that participate in protein binding without complementary enzymology and domain modeling. We propose to develop a high throughput CE-MS/MS platform for GAG structural characterization that will enable widespread engagement from the glycoscience community by incorporating CE for GAG oligosaccharide separation, a novel CE-MS interface technology, and robust tandem mass spectrometry approaches.

项目摘要/摘要 硫酸化的糖胺聚糖（GAG）碳水化合物代表了最多样化的群 生物分子和对其生物结构功能关系的全面理解尚未 实现。与其他生物分子不同，例如基于A的DNA/RNA和蛋白质 模板，堵嘴生物合成是一系列生物合成酶的累积作用的结果 产生动态的多分散混合物。该混合物的组成取决于诸如 生物年龄，发育或疾病状态以及起源组织。尽管这种多样性表现出令人生畏的 分析挑战，通过试图隔离和 表征从完整的多糖到酶形成的寡糖和 二糖。最广泛的方法是通过分离来概述脱糖液（HPLC， UHPLC，HILIC，CE）和检测（UV，荧光，质谱法（MS））。域可以 通过将这些技术合并为连字符方法来表征结构功能研究（例如LC- 多发性硬化症）。即使二糖分析是用于堵嘴分析的最广泛使用的方法，但它也无法 提供参与蛋白质结合而无需互补酶学的插科打结构基序 和域建模。我们建议开发一个高吞吐量CE-MS/MS平台，用于堵嘴结构 表征将通过合并来使聚糖社区的广泛参与 GAG寡糖分离的CE，一种新型的CE-MS接口技术和强大的串联质量 光谱法接近。