Metabolome of non-replicating M. tuberculosis

非复制结核分枝杆菌的代谢组

• 批准号: 6667206
• 负责人: Guido F Pauli
• 金额: $ 23.38万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-30 至 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6667206
• 关键词: Mycobacterium bovis; Mycobacterium tuberculosis; bacteria characteristic; bacterial genetics; cell components; cell cycle; cell wall; chromatography; drug discovery /isolation; gene environment interaction; latent bacterial disease; mass spectrometry; microorganism culture; microorganism metabolism; molecular weight; nuclear magnetic resonance spectroscopy; oxygen; tuberculosis

DESCRIPTION (provided by applicant): It is now generally accepted that new drugs are needed to make global tuberculosis control a reality. The completion of the sequence of the M. tuberculosis genome has laid the foundation for greatly accelerating new drug discovery. While a limited number of studies of the TB transcriptome and proteome have been reported, these global macromolecular analyses are still not able to contribute directly to drug discovery except where gene/protein function are known or protein crystal structures are available. Elucidating the full complement of low molecular weight compounds in an organism -the metabolome - is becoming recognized as a complementary and perhaps the most tractable approach to a comprehensive understanding of any pathogen including the insight necessary for rational drug discovery. This R-21 proposal seeks to define the metabolome of the tubercle bacillus in the stage of non-replicating persistence (NRP), the physiological state considered to be responsible for the required long treatment duration for TB. The metabolome will be elucidated by high-resolution chromatography (CCC, LC) and -spectroscopy (MS, NMR). Initial studies will utilize a high biomass of BCG to evaluate several separation schemes in optimizing the resolution of (secondary) metabolites. This protocol will then be used to define the metabolome of M. tuberculosis in NRP. Subsequent experiments will focus on understanding the growth-phase specificity of unique metabolites produced under NRP. The information obtained will not only complement the extensive knowledge of the chemistry of the cell wall but is also expected to I) identify novel secondary metabolites possibly related to those recently described for other mycobacteria, 2) help to clarify events and identify low MW markers during the metabolic shift to a low oxygen environment especially considering that >70% of genes up-regulated during this adaptation are of unknown function, 3) provide leads for drug development through analoging around TB-specific compounds not known to occur in mammalian cells and 4) assist in the understanding of the mechanism of action of newly discovered anti-TB agents. The operating hypothesis for this study is that elucidating the metabolome of Mycobacterium tuberculosis will overcome limitations inherent in global macromolecular analyses with respect to gaining key insights into dormancy and drug discovery.

描述（由申请人提供）：现在人们普遍认为需要新药才能使全球结核病控制成为现实。结核分枝杆菌基因组序列的完成为大大加速新药奠定了基础。尽管已经报道了有限的TB转录组和蛋白质组研究的研究，但除了已知基因/蛋白质功能或蛋白质晶体结构可获得基因/蛋白质功能之外，这些全球大分子分析仍无法直接贡献对药物的发现。阐明生物体中低分子量化合物的完整补体（代谢组）被认为是一种互补的，也许是对任何病原体的全面理解，包括理性药物发现所必需的见解。该R-21提案旨在在非复制持久性（NRP）的阶段定义结核菌的代谢组，该阶段被认为是TB所需的长期治疗时间的生理状态。代谢组将通过高分辨率色谱法（CCC，LC）和-Spectroscopicy（MS，NMR）阐明。初步研究将利用BCG的高生物量来评估几种分离方案，以优化（次级）代谢产物的分辨率。然后，该方案将用于定义NRP中结核分枝杆菌的代谢组。随后的实验将集中于理解NRP下产生的独特代谢产物的生长相特异性。获得的信息不仅会补充细胞壁化学的广泛知识，还可以预期i）确定新型的次级代谢产物可能与最近描述的其他分枝杆菌所描述的新型代谢物相关，2）有助于澄清事件并确定较低的MW标记并在代谢环境中转移到低氧气环境中，尤其是在该基因> 70％的情况下，在较低的氧气环境中提供了> 70％的适用于综合型号的综合型，该序列是未知的，该作用是未知的，该作用是无效的，该作用是无效的，该作用是无效的。 TB特异性化合物在哺乳动物细胞中发生，4）有助于理解新发现的抗TB剂的作用机理。这项研究的操作假设是，阐明结核分枝杆菌的代谢组将克服全球大分子分析中固有的局限性，以获得对休眠和药物发现的关键见解。