Characterization of Pupylation in Mycobacterium tuberculosis

结核分枝杆菌化脓的特征

• 批准号: 8389647
• 负责人: Katerina Heran Darwin
• 金额: $ 35.74万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-12-01 至 2015-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8389647
• 关键词: Antibiotics; Antitubercular Agents; Bacteria; Biochemical; Biochemistry; Biological; Biology; Cause of Death; Complex; Coupled; Data; Development; Drug resistance; Enzymatic Biochemistry; Enzymes; Eukaryota; Extreme drug resistant tuberculosis; Foundations; Future; Genetic; Genus Mycobacterium; Glutamates; Glutamine; Goals; Growth; In Vitro; Ligase; Ligation; Lysine; Mediating; Modeling; Molecular; Molecular Chaperones; Mus; Mycobacterium tuberculosis; Nitric Oxide; Pathogenesis; Pathway interactions; Post-Translational Protein Processing; Process; Production; Prokaryotic Cells; Proteins; Resistance; Shapes; Specificity; Structure; System; Techniques; Toxic effect; Tuberculosis; Ubiquitin; Ubiquitin Like Proteins; Virulence; base; deamidation; drug development; factor A; interest; killings; macrophage; multicatalytic endopeptidase complex; mutant; mycobacterial; novel; protein degradation; public health relevance; pup; tuberculosis drugs; tuberculosis treatment; ubiquitin ligase

DESCRIPTION (provided by applicant): Tuberculosis kills about 2 million people globally every year. A key defense against Mycobacterium tuberculosis (Mtb) infections is the production of nitric oxide (NO) by macrophages. Although NO controls Mtb growth, it rarely sterilizes the bacterium from the host, suggesting Mtb has mechanisms to resist NO toxicity. The Mtb proteasome is one such mechanism that is required for resistance to NO as well as causing death in mice. Thus, we are interested in targeting the proteasome and its associated factors for drug development. The proteasome is a multi-subunit, barrel shaped complex that degrades proteins. We found that the proteins Mpa and PafA are required for protein degradation: Mpa is thought to chaperone proteins into the proteasome core and PafA appears to be required for the attachment of a prokaryotic ubiquitin-like protein (Pup) onto substrates targeted for destruction. Pup represents the first known post-translational small protein modifier identified in any prokaryote. Little is known about how Pup is conjugated to its target substrates thus we propose to identify and characterize all proteins required for "pupylation" using genetic, biochemical and molecular biological techniques. In addition, we have recently discovered pupylation is reversible, thus we are in the process of characterizing the "depupylation" pathway. The elucidation of the Pup-proteasome sytem of Mtb will hopefully lay the foundation for the discovery and characterization of other posttranslational modification systems in all bacteria. Furthermore, these enzymes may represent new targets for the development of anti- tuberculosis drugs.

描述（由申请人提供）：每年全球约有200万人杀死约200万人。针对结核分枝杆菌（MTB）感染的关键防​​御是巨噬细胞生产一氧化氮（NO）。尽管没有对照MTB的生长，但它很少对宿主的细菌进行消毒，这表明MTB具有抵抗毒性的机制。 MTB蛋白酶体是一种这样的机制，它是对NO耐药性以及在小鼠中导致死亡所必需的一种机制。因此，我们有兴趣针对蛋白酶体及其与药物开发相关的因素。蛋白酶体是一种多纯料的桶形复合物，可降解蛋白质。我们发现，蛋白质MPA和PAFA是蛋白质降解所必需的：MPA被认为是将蛋白质蛋白蛋白与蛋白酶体核心中的伴侣蛋白，而PAFA似乎是将原核生物泛素样蛋白（PUP）连接到靶标而靶向破坏的蛋白质样蛋白（PUP）所必需的。 PUP代表了在任何原核生物中鉴定出的第一个已知的翻译后小蛋白质修饰剂。关于如何将幼崽偶联到其靶标底物，因此我们建议使用遗传，生化和分子生物学技术来识别和表征“硫化型”所需的所有蛋白质。此外，我们最近发现硫匹贝化是可逆的，因此我们正在表征“甲基化”途径。 MTB的幼崽 - 凝血体系统的阐明将为所有细菌中其他翻译后修饰系统的发现和表征奠定基础。此外，这些酶可能代表开发抗结核药物的新靶标。