Methods for Isotope Labelling of Glycans

聚糖的同位素标记方法

• 批准号: 10511205
• 负责人: Maciej Walczak
• 金额: $ 23.08万
• 依托单位: UNIVERSITY OF COLORADO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10511205
• 关键词: Address; Aldehydes; Alkylation; Amination; Amines; Area; Benchmarking; Biochemical; Carbohydrates; Carbon; Carbon Monoxide; Chemicals; Chemistry; Collection; Complement; Complex; Cytolysis; Data; Diagnosis; Disaccharides; Disease; Drug Design; Enzymatic Biochemistry; Equilibrium; Glycoconjugates; Glycoproteins; Heparan Sulfate Proteoglycan; Heparin Lyase; Heparitin Sulfate; Human; Human Milk; Interruption; Isotope Labeling; Isotopes; Kinetics; Knowledge; Label; Lead; Ligands; Link; Mass Spectrum Analysis; Mediating; Metabolic; Methodology; Methods; Modernization; Monosaccharides; Oligosaccharides; Pathologic Processes; Phenotype; Physiological Processes; Play; Polysaccharides; Positioning Attribute; Preparation; Process; Production; Property; Radiolabeled; Reaction; Role; Sampling; Sialic Acids; Source; Structure; Sulfate; System; Techniques; Technology; Therapeutic; TimeLine; Transition Elements; analog; base; biological systems; carbohydrate structure; carbonyl group; catalyst; cost; feeding; hydroxyl group; imaging study; in vivo; metabolic engineering; new technology; novel; stable isotope; sugar; technology development; tool

PROJECT SUMMARY. Glycomics studies are crucial for correlating changes in glycan structure and abundance to the phenotype of a biological system. Among the available platforms to facilitate glycomics analyses, labelling with stable isotopes has been implemented to decipher structural and quantitative aspects of carbohydrates in healthy and diseased states. Isotopically labelled saccharides are also a useful tool for in vivo metabolic profiling and in mechanistic studies in chemistry and enzymology. While some structures are commercially available, a generally applicable and non-destructive method for introducing stable heavy isotopes either in simple monosaccharides or complex oligosaccharides are not available. To overcome these limitations, we will develop a chemical technology based on carbon-13 labelling. To achieve this objective, we will pursue two specific aims: In Aim 1, we will optimize catalytic methods for selective labelling of monosaccharides with carbon-13. This method will selectively exchange the anomeric position with a heavy isotope from readily available sources of carbon-13. In Aim 2, we will apply isotope labelling method to three classes of saccharides of biomedical relevance, namely heparan sulfate proteoglycans, N-linked glycoproteins, and human milk oligosaccharides. Per requirements of PAR-19-254, we will benchmark the new technologies against the current state-of-the-art methods for oligosaccharide quantification. Upon completion of this project, a new catalytic technology for labelling of carbohydrates and glycoconjugates will be available and will complement the current tools for quantitative glycomics. In a broader sense, integration of these novel chemistries with modern mass spectrometry techniques will provide better tools for therapeutic diagnosis.

项目摘要。糖研究研究对于将聚糖结构的变化相关联至关重要 到生物系统的表型。在可用的平台中促进糖分析，标签 使用稳定的同位素已实施，以破译碳水化合物的结构和定量方面 健康和患病的状态。同位素标记的糖也是体内代谢分析的有用工具 以及化学和酶学的机械研究。虽然某些结构可商购，但 通常适用且无损的方法，用于简单地引入稳定的重型同位素 没有单糖或复杂的寡糖。为了克服这些限制，我们将发展 一种基于碳13标签的化学技术。为了实现这一目标，我们将追求两个具体的目标： 在AIM 1中，我们将优化用碳13选择性标记单糖的催化方法。这 方法将从易于使用的来源选择性地将较重的同位素交换 碳13。在AIM 2中，我们将将同位素标记方法应用于生物医学的三类糖水 相关性，即硫酸乙酰肝素蛋白聚糖，N连接的糖蛋白和人乳寡糖。每 Par-19-254的要求，我们将针对当前最新技术进行基准测试新技术 寡糖定量的方法。该项目完成后，一种新的催化技术 将提供碳水化合物和糖缀合物的标签，并将补充当前的工具 定量糖化。从广义上讲，这些新颖的化学成分与现代质量的整合 光谱技术将为治疗性诊断提供更好的工具。