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.

项目总结。糖组学研究对于将糖链结构和丰度的变化联系起来至关重要。 生物系统的表型。在现有的促进糖组分分析、标记的平台中 用稳定的同位素来破译碳水化合物的结构和数量方面 健康和患病的状态。同位素标记的糖也是体内代谢图谱的有用工具 以及化学和酶学方面的机械学研究。虽然有些结构在商业上是可用的，但 普遍适用的、非破坏性的稳定重同位素的引入方法 不提供单糖或复合低聚糖。为了克服这些限制，我们将开发 一种基于碳13标记的化学技术。为了达到这一目标，我们将追求两个具体目标： 在目标1中，我们将优化碳-13选择性标记单糖的催化方法。这 方法将从容易获得的来源中选择性地将异常体位置与重同位素交换 碳13。在目标2中，我们将把同位素标记方法应用于生物医学中的三类糖类。 相关性，即硫酸乙酰肝素蛋白多糖、N-连接糖蛋白和人乳低聚糖。人均 根据PAR-19-254的要求，我们将对照当前最先进的技术对新技术进行基准测试 低聚糖的定量方法。该项目建成后，一项新的催化技术将用于 碳水化合物和糖偶联物的标签将可用，并将补充目前的工具 定量的糖组分。在更广泛的意义上，这些新的化学物质与现代质量的结合 光谱技术将为治疗诊断提供更好的工具。