An Automated Platform for the CE-MS Analysis of Glycosaminoglycans

用于糖胺聚糖 CE-MS 分析的自动化平台

• 批准号: 10005264
• 负责人: I JONATHAN AMSTER
• 金额: $ 29.47万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10005264
• 关键词: Address; Anabolism; Animals; Automation; Binding; Binding Proteins; Biochemical Pathway; Biological; Biological Assay; Biological Process; Biology; Biopolymers; Capillary Electrophoresis; Carbohydrates; Cells; Complex; Complex Mixtures; Data; Deacetylation; Development; Disaccharides; Disease; Environment; Enzymes; Funding; GAG Gene; Generations; Glucuronic Acids; Glycosaminoglycans; Goals; Golgi Apparatus; Health; Hexosamines; Human; Infection; Ions; Laboratories; Length; Mammalian Cell; Mass Spectrum Analysis; Methods; Modification; Names; Normal Range; Organism; Outcome; Pathologic Processes; Pattern; Peptides; Physiological; Play; Polysaccharides; Procedures; Process; Protein Region; Proteins; Reagent; Regulation; Research Personnel; Role; Sampling; Series; Signal Transduction; Site; Site-Directed Mutagenesis; Specificity; Stretching; Structure; Sulfate; Surface; System; Technology; Testing; Uronic Acids; Virus; cell motility; copolymer; epimerization; innovation; instrument; interest; mass spectrometer; method development; polysulfated glycosaminoglycan; software development; tandem mass spectrometry; tool

Project Summary/Abstract    This project will develop an automated capillary electrophoresis-­tandem mass spectrometry platform for the  structural analysis of mixtures of glycosaminoglycan oligomers that are isolated in protein binding assays. This  class of carbohydrates can be found on the surface of all animal cells, and play a key role in many important  biological processes. Glycosaminoglycans are linear, polysulfated copolymers, typically with a repeating  disaccharide unit composed of an N-­acetyl-­hexosamine and glucuronic acid, and can be more than 200 repeat  units long. Modifications can occur anywhere along the chain, and these include O-­sulfation, N-­ deacetylation/N-­sulfation, and epimerization of the uronic acid. These are a particularly challenging class of  compounds to characterize due to their heterogeneous structures, an outcome of their non-­template  biosynthesis. Nevertheless, within the long glycan chains are short regions of defined modification that target  specific proteins with remarkable specificity. These glycan-­protein interactions are the basis for the biological  activity of the glycosaminoglycan. There is a critical need for tools to find these organized regions with the long  glycan chain, and to determine their pattern of modification. A few specialized laboratories have developed  mass spectrometry methods for the structural analysis of glycosaminoglycans, but these have not found their  way into biological laboratories at large. The goal of this project is to produce a platform with a high degree of  automation so that it can be deployed in the laboratories of biology researchers. The accessibility of these  advanced mass spectrometry methods will result from innovations in hardware, software, and methods  development.

项目总结/文摘

项目成果

Elucidating the Unusual Reaction Kinetics of D-Glucuronyl C5-Epimerase.

• DOI: 10.1093/glycob/cwaa035
• 发表时间: 2020-04
• 期刊: Glycobiology
• 影响因子: 4.3
• 作者: Deepika Vaidyanathan;E. Paskaleva;Troy Vargason;Xia Ke;S. McCallum;R. Linhardt;J. Dordick

A Revised Structure for the Glycolipid Terminus of Escherichia coli K5 Heparosan Capsular Polysaccharide.

• DOI: 10.3390/biom10111516
• 发表时间: 2020-11-06
• 期刊: Biomolecules
• 影响因子: 5.5
• 作者: Yan L;Fu L;Xia K;Chen S;Zhang F;Dordick JS;Linhardt RJ
• 通讯作者: Linhardt RJ