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）：结核病每年在全球造成约200万人死亡。针对结核分枝杆菌（Mtb）感染的关键防御是由巨噬细胞产生一氧化氮（NO）。虽然NO控制Mtb的生长，但它很少从宿主中杀灭细菌，这表明Mtb具有抵抗NO毒性的机制。Mtb蛋白酶体是一种这样的机制，它是抵抗NO以及导致小鼠死亡所必需的。因此，我们对靶向蛋白酶体及其相关因子的药物开发感兴趣。蛋白酶体是一种多亚基的桶形复合物，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被认为是蛋白质伴侣蛋白进入蛋白酶体核心，PafA似乎是将原核泛素样蛋白（Pup）附着到靶向破坏的底物上所必需的。Pup代表了在任何原核生物中鉴定的第一个已知的翻译后小蛋白修饰物。关于Pup如何与其靶底物缀合知之甚少，因此我们建议使用遗传，生物化学和分子生物学技术鉴定和表征“pupylation”所需的所有蛋白质。此外，我们最近发现，pupylation是可逆的，因此，我们正在表征的过程中的“去pupylation”途径。结核分枝杆菌Pu蛋白酶体系统的阐明将有望为发现和鉴定所有细菌中的其他翻译后修饰系统奠定基础。此外，这些酶可能代表抗结核药物开发的新靶点。