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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

基本信息

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
项目类别：
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

项目摘要

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.

项目摘要广泛的糖基化覆盖了更高水平生物体的细胞外环境。这些聚糖结构附着在人类蛋白质组的一半上，并且对于它们在几乎所有蛋白质组中的功能至关重要。生物过程。监测糖脂和糖蛋白上糖基化结构的改变可以发现抑制感染的新方法，并确定检测和治疗自身免疫性疾病的新靶点，疾病和癌症。尽管它们在生物学上很重要，但我们监测大量聚糖的能力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，这提供了无与伦比的结构水平，多样性用于大规模糖组学研究的工具不能提供必要的结构细节水平，有效地解决聚糖结构中的细微差异。另一方面，技术能够详细聚糖的结构解析是缓慢的、实验密集的，并且经常需要多种方法。这一项提案旨在开发一个分析平台，该平台将实现快速聚糖表征，现有技术无法提供的结构细节。该提案旨在结合离子迁移率（IMS）和气相氢/氘交换 (HDX)用于在强大的质谱平台中进行聚糖结构分析。我们将利用这个平台，新型内标物，用于分析单糖、低聚糖、聚糖链库糖蛋白和糖脂来组装离子迁移率（Aim 1）和氢/氘的数据库交换（目标2）生物相关的碳水化合物离子的性质。分子模型将被用来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）。离子迁移率和氢/氘交换的组合将被实施为色谱时间标度“GlySECT”上聚糖结构解析的LC-MS平台（目的4）。在与此同时，我们将开发软件，用于提取数据和检索已建立的数据库在这个项目中，以帮助阐明聚糖链的结构。这项技术有可能使快速、位点特异性、聚糖结构测定，并为糖生物学、生物标志物发现和治疗性蛋白质表征。

项目成果

期刊论文数量（8）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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

DOI：
10.1021/acs.chemrev.1c00279
发表时间：
2022-04-27
期刊：
CHEMICAL REVIEWS
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

Rapid Differentiation of Chondroitin Sulfate Isomers by Gas-phase Hydrogen-deuterium Exchange.