TB LAM AND AG--STRUCTURES CORRELATED WITH BIOLOGY

TB LAM 和 AG——与生物学相关的结构

• 批准号: 6373436
• 负责人: DELPHI CHATTERJEE
• 金额: $ 18.65万
• 依托单位: COLORADO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-06-01 至 2003-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6373436
• 关键词: Mycobacterium tuberculosis; SDS polyacrylamide gel electrophoresis; T lymphocyte; bacterial antigens; carbohydrate receptor; carbohydrate structure; cell wall; cellular immunity; chemical structure function; drug resistance; galactans; macrophage; mannans; mass spectrometry; nuclear magnetic resonance spectroscopy; receptor sensitivity; surface antigens

DESCRIPTION (Adapted from the Applicant's Abstract): Important physiological and immunopathogenic features of mycobacteria including slow growth, acid fastness, resistance to drug permeation, and immunological persistence have been attributed to the presence of lipoarabinomannan (LAM) and arabinogalactan (AG) in the cell wall. This competitive renewal of Al 37139 is built on success in the initial funding period of determining structural characteristics of truncated LAM from laboratory induced ethambutol (Emb) resistant M. smegmatis and LAM and AG from clinical isolates of M. tuberculosis resistant to Emb. To take the next step in fully understanding the cell surface of M. tuberculosis and its manifestation in bacterial physiology and pathology, it is proposed to determine the entire structures of LAM and AG, to characterize LAM and AG in a panel of Emb resistant M. tuberculosis isolates, and, in collaboration, to determine and precise structures of LAM needed for CD1 T-cell and macrophage mannose receptor recognition. Although some structural features of LAM and AG have been delineated, key issues, including the structure of the arabinan regions of both polymers, and the site of the arabinan attachments to the hexose backbones of each polymer remain unknown. To obtain the complete primary structure of these two 100 glycosyl residues glycans, novel cell wall degrading enzymes will be used to prepare fragments whose structures will be determined by state-of-the-art nuclear magnetic resonance (NMR) and mass spectrometry (MS) analysis. The enzymes include seven novel arabinanases and galactofuranases secreted by a Cellulomonas species and an endo alpha-1,6 mannosidase from Bacillus circulans. NMR will focus on the analysis of fully 1 3C labeled polysaccharides and polysaccharide fragments by 2-D and 3-D techniques. Mass Spectrometry will feature a novel Q-TOF-tandem electrospray mass spectrometer to obtain sequence information. Techniques based on SDS-PAGE, endo-arabinanase cleavage, 1-D-NMR, and MS will be used to efficiently characterize LAM and AG from our panel of Emb resistant M. tuberculosis. The structural studies will directly aid the immunological studies by providing a wealth of LAM related molecules needed to decipher the structural characteristics necessary for recognition by T-cells and macrophages.

描述(改编自申请人的摘要)：重要的生理功能 分枝杆菌的免疫致病特征包括生长缓慢、酸性 耐受性、抗药物渗透性和免疫学持久性 归因于脂阿拉伯甘露聚糖(LAM)和阿拉伯半乳聚糖的存在 (AG)在细胞壁中。此次Al 37139的竞争性续订是建立在成功的基础上的 在确定结构特征的初始资金期 实验室诱导的耐乙胺丁醇(EMB)耻垢分枝杆菌的截短LAM 耐emb的结核分枝杆菌临床分离株LAM和AG。至 下一步全面了解结核分枝杆菌的细胞表面 以及它在细菌生理和病理上的表现，建议 确定LAM和AG的整个结构，以刻画LAM和AG在 结核分枝杆菌耐药菌株小组，并在合作中， CD1 T细胞和巨噬细胞所需LAM的确定和精确结构 甘露糖受体识别。虽然LAM和AG的一些结构特征 已经描述了关键问题，包括阿拉比南的结构 两种聚合物的区域，以及阿拉伯胶连接到 每种聚合物的己糖主链仍不清楚。要获得完整的初级课程，请执行以下操作 这两种100糖基残基多糖的结构、新型细胞壁降解作用 酶将被用来制备碎片，其结构将被确定 通过最先进的核磁共振(核磁共振)和质谱仪(MS) 分析。这些酶包括七种新的阿拉伯聚糖酶和半乳呋喃酶 由一种纤维单胞菌和一种内切甘露糖苷酶分泌 环状芽孢杆菌。核磁共振将集中于分析完全的13C标记 2-D和3-D技术得到的多糖和多糖片段。质量 光谱分析将采用一种新型Q-TOF串联电喷雾质谱仪 以获取序列信息。基于SDS-PAGE、阿拉伯内切酶的技术 裂解、1-D-核磁共振和MS将被用来有效地表征LAM和AG 来自我们的结核分枝杆菌耐药小组。结构研究将 通过提供丰富的LAM相关信息，直接帮助免疫学研究 分子需要破译所需的结构特征 T细胞和巨噬细胞的识别。