ANALYSIS OF GLYCOSAMINOGLYCANS USING ESI-QQQ-FTMS

使用 ESI-MS/MS-FTMS 分析糖胺聚糖

• 批准号: 7723031
• 负责人: PETER B. O'CONNOR
• 金额: $ 0.13万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-06-01 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7723031
• 关键词: Complex; Computer Retrieval of Information on Scientific Projects Database; Electrons; Funding; Glycosaminoglycans; Grant; Inorganic Sulfates; Institution; Ions; Manuscripts; Methods; Peptides; Pliability; Post-Translational Protein Processing; Protein Analysis; Proteins; Reaction; Research; Research Personnel; Resources; SHFM1 gene; Source; United States National Institutes of Health; Unspecified or Sulfate Ion Sulfates; base; instrument; interest; ionization; research study; tandem mass spectrometry

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. The new ESI-qQq-FTMS has demonstrated the ability to analyze labile post-translational modifications (PTMs) on peptides and proteins using a variety of tandem mass spectrometry methods such as Q2-CAD, SORI-CAD, and ECD. It has also been used for negative ion EDD experiments. The ability to ionize and analyze molecules containing labile PTMs and the ability to use negative ion-electron reactions to do radical-based fragmentation suggests interesting capailities for the analysis of glycosaminoglycans. Due to the lability of he sulfate moieties on glycosaminoglycans, it has generally been difficult to ionize these molecules without loss of sulfates. However, initial experiments with the ESI-qQq-FTMS have shown that ionization of intact glycosaminoglycans is not particularly difficult if one carefully tunes the front end to minimize fragmentation. Further experiments are ongoing to then apply the flexibility of this instrument to the full analysis of these complex molecules using the full array of technigues from selected isolation and accumulation to EDD, ECD, and CAD methods that have been shown to be effective in full analysis of proteins and peptides. EDD is particularly useful and shows strong, clear crossring cleavages for negative glycosaminoglycan ions. Several examples have been generated and the manuscript is being prepared.

这个子项目是许多研究子项目中的一个 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。 新的ESI-qQq-FTMS已证明能够使用各种串联质谱方法（如Q2-CAD、SORI-CAD和ECD）分析肽和蛋白质上的不稳定翻译后修饰（PTM）。 它也被用于负离子EDD实验。 对含有不稳定PTM的分子进行定性和分析的能力以及使用负离子-电子反应进行基于自由基的片段化的能力表明了用于糖胺聚糖分析的有趣的能力。 由于糖胺聚糖上硫酸根部分的不稳定性，通常难以在不损失硫酸根的情况下使这些分子变性。 然而，ESI-qQq-FTMS的初始实验表明，如果仔细调整前端以最小化片段化，则完整糖胺聚糖的电离并不特别困难。 正在进行进一步的实验，然后将该仪器的灵活性应用于使用从选定的分离和积累到EDD、ECD和CAD方法的全部技术对这些复杂分子进行全面分析，这些方法已被证明在蛋白质和肽的全面分析中是有效的。 EDD是特别有用的，并显示出强烈的，明确的交叉环切割负糖胺聚糖离子。 已经产生了几个例子，目前正在编写草稿。