TB LAM AND AG--STRUCTURES CORRELATED WITH BIOLOGY

TB LAM 和 AG——与生物学相关的结构

• 批准号: 6170035
• 负责人: DELPHI CHATTERJEE
• 金额: $ 18.1万
• 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 2002-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6170035
• 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 分枝杆菌的免疫病特征，包括缓慢生长，酸 牢度，对药物渗透和免疫持久性具有 归因于存在脂肪动物群（LAM）和阿拉伯氏菌的存在 （Ag）在细胞壁中。 AL 37139的这种竞争性更新是基于成功的 在确定结构特征的最初资助期间 从实验室诱导的乙酰丁醇（EMB）耐药的M. smegmatis截断的lam lam and ag来自结核分枝杆菌的临床分离株耐胚胎。到 采取下一步充分了解结核分枝杆菌的细胞表面 及其在细菌生理学和病理学中的表现，提出 确定LAM和AG的整个结构，以表征A中的Lam和Ag 抗胚胎抗性分枝杆菌分离株的面板，并合作 确定CD1 T细胞和巨噬细胞所需的LAM的精确结构 甘露糖受体识别。虽然Lam和Ag的某些结构特征 已经被描述了关键问题，包括阿拉伯人的结构 两种聚合物的区域以及阿拉伯人附着的地点 每种聚合物的己糖骨架仍然未知。获得完整的初级 这两个100个糖基残基聚糖的结构，新的细胞壁降解 酶将用于制备将确定结构的碎片 通过最先进的核磁共振（NMR）和质谱法（MS） 分析。这些酶包括七个新型阿拉伯酶和半乳呋喃酶 由纤维素瘤物种和一个alpha-1,6甘露糖苷酶分泌 马戏团芽孢杆菌。 NMR将重点放在完全1 3C标记的分析上 通过2-D和3-D技术，多糖和多糖片段。大量的 光谱法将具有新颖的Q-TOF串联电喷雾质谱仪 获取序列信息。基于SDS-PAGE，恩托 - 阿拉宾酶的技术 切割，1-D-NMR和MS将用于有效表征LAM和Ag 从我们的耐Emb抗性结核分枝杆菌。结构研究将 通过提供大量相关的LAM，直接帮助免疫研究 破译所需的分子所需的分子 T细胞和巨噬细胞的识别。