Hybrid structural mass spectrometry for rapid site-specific glycan structural elucidation

用于快速位点特异性聚糖结构解析的混合结构质谱法

• 批准号: 10186777
• 负责人: Miklos Guttman
• 金额: $ 42.67万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10186777
• 关键词: Adopted; Autoimmune Diseases; Behavior; Biochemistry; Biological; Biological Markers; Biological Process; Biological Products; Carbohydrates; Cells; Chromatography; Complex; Complex Mixtures; Computer software; Data; Data Set; Databases; Deuterium; Deuterium Exchange Measurement; Diagnostic; Disease; Ensure; Gases; Glycobiology; Glycoconjugates; Glycolipids; Glycopeptides; Glycoproteins; Goals; Human; Hybrids; Hydrogen; Infection; Ions; Isomerism; Kinetics; Libraries; Location; Malignant Neoplasms; Mass Spectrum Analysis; Measurement; Measures; Methods; Modification; Molecular Conformation; Monitor; Monosaccharides; Nature; Oligosaccharides; Onset of illness; Organism; Outcome; Pattern; Phase; Polymers; Polysaccharides; Preparation; Property; Proteome; Protocols documentation; Reference Standards; Reproducibility; Role; Sampling; Series; Site; Software Tools; Spectrometry; Speed; Structure; Techniques; Technology; Testing; Therapeutic Intervention; Work; base; biomarker discovery; cancer cell; carbohydrate structure; data acquisition; data exchange; extracellular; glycoproteomics; glycosylation; in silico; ion mobility; millisecond; molecular modeling; new technology; novel; pathogen; prevent; rapid technique; software development; sugar; therapeutic protein; tool

PROJECT SUMMARY An extensive array of glycosylation covers the extracellular milieu of higher level organisms. These glycan structures are attached to half of the human proteome and are critical for their function in nearly all biological processes. Monitoring alterations in the structures of glycosylation on glycolipids and glycoproteins can identify new ways of inhibiting infections and pinpoint new targets for detecting and treating autoimmune diseases and cancer. Despite their biological importance, our ability to monitor the vast array of glycan structures and their exact role in disease states remains limited. This is primarily due to the challenges associated with analyzing complex carbohydrate structures, which present an unparalleled level of structural diversity. Tools for large-scale glycomic studies do not provide the level of structural detail necessary to effectively resolve subtle differences in glycan structures. On the other hand, techniques capable of detailed structural elucidation of glycans are slow, experimentally intensive, and often require multiple approaches. This proposal seeks to develop an analytical platform that will enable rapid glycan characterization with a level of structural detail that is unavailable with existing techniques. This proposal aims to incorporate ion mobility (IMS) and gas-phase hydrogen/deuterium exchange (HDX) for glycan structural analysis into a robust mass spectrometry platform. We will use this platform with novel internal standards to analyze a library of monosaccharides, oligosaccharides, glycan chains from glycoproteins, and glycolipids to assemble a database of the ion mobility (Aim 1) and hydrogen/deuterium exchange (Aim 2) properties of biologically relevant carbohydrate ions. Molecular modeling will be used to help interpret and rationalize the different behaviors of each structure and expand our structural reference database in silico (Aim 3). The combination of ion mobility and hydrogen/deuterium exchange will be implemented into a LC-MS platform for Glycan Structural Elucidation on a Chromatographic Timescale “GlySECT” (Aim 4). In parallel, we will develop the software for extracting the data and searching against the databases established in this project to aid in structural elucidation of the glycan chains. This technology has the potential to enable rapid, site-specific, glycan structural determination and provide a much-needed tool for glycobiology, biomarker discovery, and therapeutic protein characterization.

项目概要 广泛的糖基化覆盖了高级生物体的细胞外环境。 聚糖结构附着在一半的人类蛋白质组上，对其几乎所有功能都至关重要 监测糖脂和糖蛋白糖基化结构的变化。 可以找到抑制感染的新方法并确定检测和治疗自身免疫的新靶标 尽管它们具有生物学重要性，但我们有能力监测大量的聚糖。 结构及其在疾病状态中的确切作用仍然有限，这主要是由于挑战。 与分析复杂的碳水化合物结构相关，其呈现出无与伦比的结构水平 大规模糖组学研究的工具无法提供必要的结构细节水平。 另一方面，技术能够有效地解决聚糖结构的细微差异。 聚糖的结构解析过程缓慢，实验强度大，并且通常需要多种方法。 该提案旨在开发一个分析平台，能够以一定水平快速进行聚糖表征 现有技术无法获得的结构细节。 该提案旨在结合离子淌度 (IMS) 和气相氢/氘交换 (HDX) 用于聚糖结构分析的强大质谱平台我们将使用该平台。 用于分析单糖、寡糖、聚糖链库的新颖内标 糖蛋白和糖脂来组装离子淌度（目标 1）和氢/氘的数据库 生物学相关碳水化合物离子的交换（目标 2）特性将用于帮助。 解释和合理化每个结构的不同行为，并扩展我们的结构参考数据库 计算机模拟（目标 3）将离子淌度和氢/氘交换的组合实施到计算机中。 用于在色谱时间尺度“GlySECT”上进行聚糖结构阐明的 LC-MS 平台（目标 4）。 同时，我们将开发用于提取数据并根据已建立的数据库进行搜索的软件 在这个项目中，帮助阐明聚糖链的结构。这项技术有潜力实现。 快速、位点特异性的聚糖结构测定，并为糖生物学、生物标志物提供急需的工具 发现和治疗性蛋白质表征。

项目成果

Characterization of the Impact of Drug Metabolism on the Gas-Phase Structures of Drugs Using Ion Mobility-Mass Spectrometry.

使用离子淌度质谱法表征药物代谢对药物气相结构的影响。

• DOI: 10.1021/acs.analchem.9b03292
• 发表时间: 2019-10-15
• 期刊: Analytical chemistry
• 影响因子: 7.4
• 作者: Dylan H Ross;Ryan P. Seguin;Libin Xu
• 通讯作者: Libin Xu

Linkage Memory in Underivatized Protonated Carbohydrates.

未衍生化质子化碳水化合物中的连锁记忆。

• 发表时间: 2021-02-03
• 影响因子: 3.2
• 作者: Mookherjee, Abhigya;Uppal, Sanjit S;Murphree, Taylor A;Guttman, Miklos
• 通讯作者: Guttman, Miklos

Dissection of Fragmentation Pathways in Protonated N-Acetylhexosamines.

质子化 N-乙酰己糖胺断裂途径的剖析。

• 发表时间: 2018-10-16
• 影响因子: 7.4
• 作者: Mookherjee, Abhigya;Uppal, Sanjit S;Guttman, Miklos
• 通讯作者: Guttman, Miklos

Advances in Hydrogen/Deuterium Exchange Mass Spectrometry and the Pursuit of Challenging Biological Systems.

氢/氘交换质谱的进展和对具有挑战性的生物系统的追求。

• 发表时间: 2022
• 影响因子: 62.1
• 作者: James, Ellie I;Murphree, Taylor A;Vorauer, Clint;Engen, John R;Guttman, Miklos
• 通讯作者: Guttman, Miklos

High-Precision, Gas-Phase Hydrogen/Deuterium-Exchange Kinetics by Mass Spectrometry Enabled by Exchange Standards.

通过交换标准实现的高精度气相氢/氘交换动力学质谱分析。

• 发表时间: 2020
• 影响因子: 7.4
• 作者: Uppal, Sanjit S;Mookherjee, Abhigya;Harkewicz, Rick;Beasley, Sarah E;Bush, Matthew F;Guttman, Miklos
• 通讯作者: Guttman, Miklos