TB LAM AND AG--STRUCTURES CORRELATED WITH BIOLOGY

描述（改编自申请人的摘要）：重要的生理学 和分枝杆菌的免疫病理学特征，包括生长缓慢、酸性 耐药性、耐药物渗透性和免疫持久性， 由于存在脂阿拉伯甘露聚糖（LAM）和阿拉伯半乳聚糖 (AG)在细胞壁中。Al 37139的这种竞争性更新是建立在成功的基础上的 在确定结构特征的初始供资期间， 从实验室诱导的乙胺丁醇（Emb）抗性M.耻垢 从临床分离的M.对Emb耐药的结核病到 采取下一步充分了解M的细胞表面。结核 及其在细菌生理学和病理学中的表现，建议 确定LAM和AG的整个结构，以表征LAM和AG在 Emb抗性M.结核病分离株，并与合作， 确定和精确的LAM的结构所需的CD 1 T细胞和巨噬细胞 甘露糖受体识别虽然LAM和AG的一些结构特征 已经划定，关键问题，包括阿拉伯半岛的结构， 这两种聚合物的区域，以及阿拉伯聚糖连接到聚合物的位点。 每种聚合物的己糖主链仍然是未知的。为了获得完整的初选 这两种结构的100个糖基残基聚糖，新型细胞壁降解 酶将用于制备片段，其结构将被确定 通过最先进的核磁共振（NMR）和质谱（MS） 分析.这些酶包括七种新的阿拉伯聚糖酶和半乳糖呋喃酶 由纤维单胞菌属物种分泌的内切α-1，6甘露糖苷酶， 环状芽孢杆菌核磁共振将集中在分析完全13 C标记的 多糖和多糖片段的2-D和3-D技术。质量 光谱将采用一种新型的Q-TOF串联电喷雾质谱仪 以获得序列信息。基于SDS-PAGE的技术，内切阿拉伯聚糖酶 裂解、1-D-NMR和MS将用于有效表征LAM和AG 从我们的抗Emb M的小组。结核结构研究将 通过提供大量与LAM相关的免疫学研究， 分子需要破译必要的结构特征， 被T细胞和巨噬细胞识别。

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.