ANALYSIS OF MYCOBACTERIUM ANTIGENS BY VIBRATIONALLY COOLED MALDI FTMS

通过振动冷却 MALDI FTMS 分析分枝杆菌抗原

• 批准号: 7369219
• 负责人: DAVID MOODY
• 金额: $ 0.41万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-01 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7369219
• 关键词: ANALYSIS; MYCOBACTERIUM; ANTIGENS; VIBRATIONALLY; COOLED

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. Recent studies of the Mycobacterium antigens have shown that the ligands recognized by CD1-restricted T-cells, are not peptides, but instead are lipids (e.g., mycolic acids) and glycolipids (e.g., lipoarabinomannan and phosphatidylinositol mannoside). The precise structures of the antigens presented by most CD1 proteins have not yet been reported. CD1-presented glycolipid antigens, for which structures have so far been determined in our laboratories and elsewhere, are composed of diverse building blocks linked by energetically labile bonds and often occur in nature as complex mixtures of closely related structures. The method the BUSM MS Resource has developed for direct coupling of thin layer chromatography (TLC) with vibrationally-cooled matrix-assisted laser desorption/ionization Fourier transform mass spectrometry (VC MALDI-FTMS) is now being applied to samples of mycobacterial lipids specifically produced in vivo. The structure of CD1-presented glycolipid antigens should be revealed, with respect to the number of the carbohydrate residues and their attachment points, as well as the precise arrangement of the branching and the structure of acyl chains. As a first step in implementing this project, a test of concentration/sensitivity has been performed using TLC plates loaded with glycolipid and phospholipid standards, such as GMM, DDM, and PI.

该子项目是利用 NIH/NCRR 资助的中心拨款提供的资源的众多研究子项目之一。子项目和研究者 (PI) 可能已从另一个 NIH 来源获得主要资金，因此可以在其他 CRISP 条目中得到体现。列出的机构是中心的机构，不一定是研究者的机构。最近对分枝杆菌抗原的研究表明，CD1 限制性 T 细胞识别的配体不是肽，而是脂质（例如分枝菌酸）和糖脂（例如脂阿拉伯甘露聚糖和磷脂酰肌醇甘露糖苷）。大多数 CD1 蛋白呈递的抗原的精确结构尚未报道。 CD1 呈递的糖脂抗原的结构迄今已在我们的实验室和其他地方确定，它由通过能量不稳定键连接的不同构建块组成，并且通常以密切相关结构的复杂混合物形式存在于自然界中。 BUSM MS Resource 开发的用于将薄层色谱 (TLC) 与振动冷却基质辅助激光解吸/电离傅里叶变换质谱 (VC MALDI-FTMS) 直接耦合的方法现已应用于体内专门产生的分枝杆菌脂质样品。应揭示 CD1 呈递的糖脂抗原的结构，包括碳水化合物残基的数量及其附着点，以及分支的精确排列和酰基链的结构。作为实施该项目的第一步，使用装载了糖脂和磷脂标准品（例如 GMM、DDM 和 PI）的 TLC 板进行了浓度/灵敏度测试。