Methods for Isotope Labelling of Glycans

聚糖的同位素标记方法

基本信息

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
项目类别：
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

项目摘要

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生物系统的表型。在促进糖组学分析的可用平台中，标记稳定同位素已被用来破译碳水化合物的结构和数量方面健康和患病状态。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。虽然有些结构是可商用的，普遍适用且非破坏性的方法，用于以简单的方式引入稳定的重同位素单糖或复合寡糖不可用。为了克服这些限制，我们将开发基于碳13标记的化学技术。为了实现这一目标，我们将追求两个具体目标：在目标 1 中，我们将优化用碳 13 选择性标记单糖的催化方法。这该方法将选择性地与来自容易获得的来源的重同位素交换异头位置碳13。在目标2中，我们将把同位素标记方法应用于生物医学的三类糖类相关性，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对新技术进行基准测试寡糖定量方法。该项目完成后，将采用一种新的催化技术碳水化合物和糖复合物的标记将可用，并将补充现有的工具定量糖组学。从更广泛的意义上讲，这些新颖的化学物质与现代大众的结合光谱测定技术将为治疗诊断提供更好的工具。