ELECTRON TRANSFER DISSOCIATION OF GLYCANS AND GLYCOCONJUGATES

聚糖和糖缀合物的电子转移解离

• 批准号: 8365562
• 负责人: Catherine E. Costello
• 金额: $ 5.08万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2012-08-09
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8365562
• 关键词: American; Anions; Biology; Carbohydrates; Cells; Charge; Chemicals; Data; Databases; Dissociation; Electron Transport; Electrons; Exhibits; Fourier transform ion cyclotron resonance; Funding; Gases; Glycoconjugates; Glycopeptides; Grant; Ions; Journals; Label; Laboratories; Link; Manuscripts; Mass Spectrum Analysis; Medicine; Methods; Milk; National Center for Research Resources; Pattern; Peptides; Phase; Polysaccharides; Principal Investigator; Proteins; Publishing; Reagent; Research; Research Infrastructure; Resources; SHFM1 gene; Societies; Source; United States National Institutes of Health; Ursidae Family; Variant; analog; base; cost; fluoranthene; instrument; ionization; mass spectrometer; research study

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Although ECD has proven its usefulness for proteins, peptides and carbohydrates, its applications are limited by the fact that its use is almost completely restricted to Fourier transform ion cyclotron resonance mass spectrometers. To make electron-based methods practical on instruments other than FTMS instruments, Hunt et al. introduced a new method for dissociation that relies on electron transfer by negative species (usually fluoranthene anions) that are produced in a negative-ion chemical ionization source and then brought into contact, in the gas phase, with multiply-charged positive ions that have been generated by ESI and trapped in a cell. Because of the excellent results that we had obtained by ECD and EDD of glycans, we undertook a new project to explore the utility of ETD for analysis of glycans and glycoconjugates. Since we did not yet have an instrument equipped for ETD in the Resource, we began our studies by taking advantage of an offer from Bruker Daltonics, Inc., who had an appropriately equipped quadrupole ion trap instrument available at their headquarters in nearby Billerica, MA. Our initial experiments produced spectra that have rich patterns that include both glycosidic and cross-ring cleavages. They therefore bear strong similarities to the ECD spectra of glycans that we have already obtained, but also exhibit additional fragments, and thus the data is complementary. In April 2009, Bruker installed one of their new ion trap instruments (amaZon ETD) in our laboratory, so that we may continue these studies and also generate reference tandem mass spectra for glycans that we will make available for inclusion in public databases. The mass spectra obtained by CID and ETD, alone and in combination, of a set of milk glycans have been obtained, for the native compounds, and for the permethylated derivatives of the native, reduced and O-18 labelled analogs. These were exhaustively interpreted and the resulting manuscript was published in the Journal of the American Society for Mass Spectrometry. Further analyses have included N-linked glycans and glycopeptides. In addition, variation of the chemical reagent for ETD and the chemical ionization gas is being evaluated to optimize the approach for glycans.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 尽管ECD证明了其对蛋白质，肽和碳水化合物的有用性，但其应用受到了以下事实的限制，即它的使用几乎完全局限于傅立叶转化离子循环共振共振质谱仪。为了使基于电子的方法对FTMS仪器以外的其他仪器进行实用，Hunt等人。引入了一种新的解离方法，该方法依赖于通过负化化学电离源产生的负物质（通常是氟阴离子）的电子转移，然后在气相中接触，并在气相中与ESI产生的多重阳性离子并被捕获在细胞中。由于我们由Glycans的ECD和EDD获得的良好结果，我们进行了一个新项目，以探索ETD用于分析聚糖和糖缀合物的实用性。由于我们还没有资源中的ETD配备工具，因此我们利用Bruker Daltonics，Inc。的要约开始了研究，后者在马萨诸塞州Billerica附近的总部提供了适当的四极离子陷阱仪器。我们的最初实验产生的光谱具有丰富的模式，包括糖苷和跨环切割。因此，它们与我们已经获得的聚糖的ECD光谱具有很强的相似性，但也表现出其他片段，因此数据是互补的。 2009年4月，布鲁克（Bruker）在我们的实验室中安装了一种新的离子陷阱仪器（Amazon ETD），以便我们可以继续这些研究，并为Glycans生成参考串联质谱，我们将可以将其包含在公共数据库中。对于天然化合物，已经获得了一组牛奶聚糖的CID和ETD获得的质谱，对于天然化合物，对天然的氯二甲基化衍生物，降低和O-18标记的类似物。这些都是详尽的解释，由此产生的手稿发表在《美国质谱学会杂志》上。进一步的分析包括N连接的聚糖和糖肽。另外，正在评估ETD化学试剂和化学电离气体的变化，以优化聚糖方法。