ANALYTICAL TECHNIQUES FOR CARBOHYDRATE STRUCTURE DETERMINATION

碳水化合物结构测定的分析技术

• 批准号: 7956012
• 负责人: Parastoo Azadi
• 金额: $ 0.13万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-01 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7956012
• 关键词: Acetates; Amber; Area; Biochemical; Carbohydrate Conformation; Carbohydrates; Computer Retrieval of Information on Scientific Projects Database; Computer Simulation; Digestion; Exoglycosidases; Funding; Glycoconjugates; Glycoproteins; Grant; Hand; High Pressure Liquid Chromatography; Information Resources; Institution; Laboratories; Literature; Mass Fragmentography; Mass Spectrum Analysis; Methods; Methylation; Monosaccharides; NMR Spectroscopy; Oligosaccharides; Participant; Polysaccharides; Production; Protocols documentation; Research; Research Personnel; Resources; Sampling; Site; Source; Structure; Sugar Alcohols; Techniques; Technology; United States National Institutes of Health; Validation; Work; carbohydrate structure; chemical synthesis; experience; glycosylation; instrument; lectures; molecular dynamics; preference; sugar

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Analytical Techniques for Carbohydrate Structural Determination (one-week course) This is a one-week course which covers the techniques of glycosyl-residue and glycosyl-linkage composition analyses using gas-liquid chromatography/mass spectrometry (GC/MS). The hands-on work includes the derivatization of sugar samples to make alditol acetates and trimethylsilyl derivatives and the production of partially methylated alditol acetates that the trainees analyze by GC/MS. Lectures and demonstrations cover such techniques for structural analysis MALDI-MS ans ESI-MS mass spectrometry, ^1 H-NMR spectroscopy, and methods for the separation and purification of monosaccharides and oligosaccharides using Metrohm peak HPLC and other chromatographic techniques. Additional lectures are also given on specific topics such as Chemical synthesis of oligosaccharides and isolation and purification of polysaccharides This course covers the techniques of glycosyl residue (composition) and glycosyl linkage (methylation) analyses using gas-liquid chromatography/mass spectrometry. Participants perform analyses of purified oligosaccharides, and they may bring one of their own samples. Lectures and demonstrations cover other techniques for structural analysis. In addition, a choice of modules either on using mass spectrometry or NMR spectroscopy for analysis of glycoconjugates is included (see descriptions below). Experience with basic biochemical techniques is a prerequisite for participation. Additional Course Modules Mass Spectrometric (MS) Analysis of Glycoconjugates: The use of mass spectrometry for the characterization of glycoconjugates is discussed. Topics in this area include identifying occupied glycosylation sites by LC-MS and sequencing glycoprotein glycans with sequential exoglycosidase digestions followed by MS. Laboratory demonstrations are performed on these two topics. NMR of Carbohydrates: The NMR module consists of an introduction to the application of NMR spectroscopy to polysaccharides and glycoconjugates covering (i) essential features of NMR spectra, (ii) protocols used to solve primary structures, (iii) applications and examples from the current literature, and (iv) resources for information. Our four NMR instruments (300-, 500-, 600-, and 800-MHz) are used for demonstrations during this module. Computational Simulations of Carbohydrate Conformation: The course presents a discussion of the conformational preferences of oligosaccharides. Focus shifts to the prediction and validation of oligosaccharide structures produced computationally with molecular dynamics simulations with the AMBER/GLYCAM force field.

这个子项目是许多利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 碳水化合物结构测定的分析技术（为期一周的课程） 这是一个为期一周的课程，涵盖了使用气液色谱/质谱（GC/MS）进行糖基残基和糖基键组成分析的技术。 实践工作包括糖样品的衍生化以制备糖醇乙酸酯和三甲基甲硅烷基衍生物，以及学员通过GC/MS分析的部分甲基化糖醇乙酸酯的生产。讲座和演示涵盖了结构分析MALDI-MS和ESI-MS质谱，^1H-NMR光谱，以及使用瑞士万通峰HPLC和其它色谱技术分离和纯化单糖和寡糖的方法。 另外还提供了关于特定主题的讲座，如寡糖的化学合成和多糖的分离和纯化 本课程涵盖了糖基残基（组成）和糖基连接（甲基化）的分析技术，使用气液色谱/质谱。 参与者对纯化的低聚糖进行分析，他们可以携带自己的样品。 讲座和演示涵盖了结构的其他技术 分析. 此外，还包括使用质谱法或NMR光谱法分析糖缀合物的模块选择（见下文描述）。 具有基本生物化学技术的经验是参加的先决条件。 附加课程模块 糖缀合物的质谱（MS）分析：讨论了糖缀合物表征的质谱法的使用。 在这方面的主题包括确定占领的糖基化位点的LC-MS和测序糖蛋白聚糖与顺序外切糖苷酶digestion随后由MS。实验室演示进行这两个主题。 碳水化合物的NMR：NMR模块包括介绍NMR光谱在多糖和糖缀合物中的应用，涵盖（i）NMR光谱的基本特征，（ii）用于解决一级结构的协议，（iii）当前文献中的应用和示例，以及（iv）信息资源。 我们的四台NMR仪器（300-，500-，600-和800-MHz）用于本模块的演示。 碳水化合物构象的计算机模拟：本课程讨论寡糖的构象偏好。 重点转移到预测和验证寡糖结构的计算与分子动力学模拟与琥珀/GLYCAM力场。