Characterization of the nutrient assimilation pathways in M. tuberculosis

结核分枝杆菌营养同化途径的特征

基本信息

批准号：
10304930
负责人：
Brian C VanderVen
金额：
$ 64.02万
依托单位：
CORNELL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-11-19 至 2025-10-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10304930
关键词：
ATP phosphohydrolase Anti-Inflammatory Agents Arachidonic Acids Assimilations Bacteria Biochemical Biochemical Pathway Carbon Cessation of life Characteristics Chemicals Cholesterol Chronic Complex Data Degradation Pathway Dinoprostone Disease Drug Targeting Eicosanoids Enzymes Fatty Acids Funding Gene Proteins Genetic Granuloma Human Immune Immune response Immune signaling Infection Knowledge Light Lipids Lung Mediating Metabolic Metabolic Pathway Modeling Mus Mycobacterium tuberculosis Nutrient Operon Pathogenesis Pathway interactions Pharmaceutical Preparations Phenotype Play Positioning Attribute Process Protein Subunits Proteins Research Role Source Starvation Substrate Specificity System Therapeutic Time Tuberculosis Virulence Virulence Factors Work bacterial fitness base chronic infection drug development fatty acid transport genetic approach immunoregulation in vivo macrophage mouse model mutant novel nutrition pathogen pathogenic bacteria

项目摘要

Project Summary / Abstract Mtb utilizes host-derived lipids to promote pathogenesis and this is a defining feature of this intracellular pathogen. During infection Mtb imports and metabolizes host lipids to support pathogenesis by producing: i) energy, ii) central metabolic intermediates, or iii) polyketide virulence lipids. While the metabolic pathways in Mtb that degrade or process lipids are complex and contain redundant enzymes, the bacterial Mce lipid transporters appear to be specific for dedicated lipid substrates. Aim 1 of this work proposes to employ genetic and biochemical approaches to identify and characterize novel gene/proteins required for fatty acid import in Mtb. While it is understood that Mce1 imports fatty acids, the substrate specificity of this transporter is unknown. Therefore, we intend to define substrates and the biochemical basis of Mce1 substrate specificity. Our preliminary studies indicate that Mtb transports fatty acid precursors of immune signaling lipids via Mce1 and we include here studies to evaluate if scavenging of this immune lipid precursors by Mtb impacts the immune response. Aim 2 proposes to identify and characterize protein subunits that are shared by all the Mce transporters and are required for lipid import in Mtb. We have determined that LucA is required for Mce1- and Mce4-mediated transport and LucA stabilizes these transporter complexes. These studies seek to characterize the basis for this transporter stabilization. Similarly, MceG is required for Mce1- and Mce4-mediated transport and we intend to understand how MceG stabilizes and interacts with Mce1. We will use a genetic approach to silence LucA and MceG in Mtb within chronically infected mice and quantify bacterial fitness to determine the therapeutic potential of drugs that potentially block these proteins.

项目摘要 /摘要 MTB利用宿主衍生的脂质来促进发病机理，这是该细胞内的定义特征病原。在感染期间，MTB进口并代谢宿主脂质以通过产生：I）来支持发病机理能量，ii）中央代谢中间体或iii）聚酮化合物毒力脂质。而MTB的代谢途径降解或过程脂质是复杂的，含有冗余酶，细菌MCE脂质转运蛋白似乎针对专用的脂质底物。这项工作的目的1建议采用遗传和生化方法来识别和表征新颖脂肪酸在MTB中进口所需的基因/蛋白质。虽然可以理解MCE1进口脂肪酸，但该转运蛋白的底物特异性未知。因此，我们打算定义底物和生化 MCE1底物特异性的基础。我们的初步研究表明，MTB转运的脂肪酸前体通过MCE1进行免疫信号脂质，我们在这里包括研究该免疫脂质是否清除 MTB的前体会影响免疫反应。 AIM 2建议识别和表征所有MCE转运蛋白共享的蛋白质亚基和在MTB中导入脂质所需。我们已经确定MCE1和MCE4介导的LUCA是必需的运输和卢卡稳定这些转运蛋白络合物。这些研究旨在为此奠定基础转运蛋白稳定。同样，MCE1和MCE4介导的运输需要MCEG，我们打算了解MCEG如何稳定并与MCE1相互作用。我们将使用一种遗传方法来沉默卢卡和 MCEG在慢性感染小鼠中MTB中的MCEG并量化细菌适应性以确定治疗潜力可能阻止这些蛋白质的药物。