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。糖组学研究对于关联聚糖结构和丰度的变化至关重要 一个生物系统的表型。在促进糖组学分析的可用平台中，标记 已经实施了稳定同位素来破译碳水化合物的结构和定量方面， 健康和疾病状态。同位素标记的同位素标记物也是体内代谢谱分析的有用工具 以及化学和酶学的机理研究。虽然一些结构是商业上可获得的，但是 用于引入稳定的重同位素的普遍适用的和非破坏性的方法 单糖或复合低聚糖不可用。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。为达致这个目标，我们会致力达致两个具体目标： 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 该方法将选择性地用来自容易获得的重同位素来源的异头位置交换。 碳13目标二：将同位素标记方法应用于三类生物医学药物的研究， 相关性，即硫酸乙酰肝素蛋白聚糖、N-连接糖蛋白和人乳寡糖。每 根据PAR-19-254的要求，我们将根据当前最先进的技术水平对新技术进行基准测试 寡糖定量的方法。在该项目完成后，一种新的催化技术， 碳水化合物和糖缀合物的标记将是可用的，并将补充目前的工具， 定量糖组学从更广泛的意义上说，将这些新的化学物质与现代物质相结合， 光谱技术将为治疗性诊断提供更好的工具。