Functional phosphoinositide lipidomics of Legionella-containing vacuoles

含军团菌液泡的功能性磷酸肌醇脂质组学

• 批准号: 198140865
• 负责人: Professor Dr. Hubert Hilbi
• 金额: --
• 依托单位: Institut für Medizinische Mikrobiologie
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2011
• 资助国家: 德国
• 起止时间: 2010-12-31 至 2014-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/198140865?language=en
• 关键词: Functional; phosphoinositide; lipidomics; Legionella; containing

Legionella spp. are environmental bacteria that grow within free-living amoebae and within macrophages in the lung, thereby causing a severe pneumonia termed “Legionnaires’ disease”. The bacteria replicate in host cells in distinct membrane-bound compartments, the “Legionella-containing vacuoles” (LCVs). Formation of LCVs is a complex and robust process, which involves a bacterial type IV secretion system and more than 100 different secreted “effector” proteins. These proteins are injected into host cells, where they modulate signal transduction and membrane dynamics. The aims of this project comprise a detailed molecular analysis using biochemical, genetic and cellular microbial methods of host phosphoinositide (PI) lipids and other lipids implicated in LCV formation by Legionella spp. To this end, we will purify intact LCVs and determine the PIs, the lipidome and the proteome by mass spectrometry. To functionally characterize the role of PIs in the formation of LCVs, we will (i) establish and compare PI and lipid catalogs of purified intact LCVs of Legionella pneumophila and Legionella longbeachae, (ii) analyze the dynamics of PIs and PImetabolizing enzymes on LCVs in real-time, (iii) characterize PI kinases and PI phosphatases implicated in LCV formation, and (iv) study the functional role of PIs for LCV formation.

军团菌属是在自由生活的变形虫和肺部巨噬细胞内生长的环境细菌，从而引起称为“军团病”的严重肺炎。细菌在宿主细胞中不同的膜结合区室（“含军团菌液泡”（LCV））中复制。 LCV 的形成是一个复杂而稳健的过程，涉及 IV 型细菌分泌系统和 100 多种不同的分泌“效应”蛋白。这些蛋白质被注射到宿主细胞中，调节信号转导和膜动力学。该项目的目的包括使用生化、遗传和细胞微生物方法对宿主磷酸肌醇 (PI) 脂质和其他与军团菌 LCV 形成有关的脂质进行详细的分子分析。为此，我们将纯化完整的 LCV，并通过质谱法测定 PI、脂质组和蛋白质组。为了从功能上表征 PI 在 LCV 形成中的作用，我们将 (i) 建立并比较嗜肺军团菌和长滩军团菌纯化的完整 LCV 的 PI 和脂质目录，(ii) 实时分析 LCV 上 PI 和 PI 代谢酶的动态，(iii) 表征相关的 PI 激酶和 PI 磷酸酶 (iv) 研究 PI 对 LCV 形成的功能作用。