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.

项目总结/摘要 硫酸化糖胺聚糖（GAG）碳水化合物代表了结构上最多样化的糖胺聚糖类之一。 生物分子及其生物结构-功能关系的全面理解还有待于 实现。不像其他生物分子，如DNA/RNA和蛋白质，是基于一个合成的。 模板，GAG生物合成是一系列生物合成酶累积作用的结果， 产生动态的多分散混合物。该混合物的组成取决于以下因素， 生物体年龄、发育或疾病状态以及来源组织。尽管这种多样性呈现出令人生畏的 分析方面的挑战，通过尝试分离和 表征从完整多糖到酶促制备的寡糖的GAG， 二糖最普遍的方法是通过分离（HPLC， UHPLC、HILIC、CE）和检测（UV、荧光、质谱（MS））。结构域可以 通过将这些技术结合到联用方法（例如LC-120）中来表征结构-功能研究。 MS）。尽管二糖分析是最广泛使用的GAG分析方法，但它不能用于分析。 在GAG中提供参与蛋白质结合的结构基序，而无需互补酶学 域建模。我们建议开发一个高通量的毛细管电泳-串联质谱（CE-MS/MS）平台，用于GAG结构分析。 表征，这将使糖科学界的广泛参与， 用于GAG寡糖分离的CE，一种新型CE-MS接口技术和稳健的串联质谱 光谱方法。