The analysis of membrane signalling networks in latex bead- and mycobacterial-phagosomes regulating actin binding and de novo assembly

乳胶珠和分枝杆菌吞噬体中调节肌动蛋白结合和从头组装的膜信号网络分析

• 批准号: 5407324
• 负责人: Professor Dr. Thomas Dandekar
• 金额: --
• 依托单位: Institut für Biowissenschaften (IfBi)
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2003
• 资助国家: 德国
• 起止时间: 2002-12-31 至 2007-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/5407324?language=en
• 关键词: analysis; membrane; signalling; networks; latex

Two in vitro assays have been developed to analyse how a defined cytoplasmic membrane surface, latex bead phagosomes (LBP) from macrophages (A) assembles actin de novo and (B) binds to F-actin. A large network of proteins and lipids has been identified that regulates the assembly process. The phagosomes of non-pathogenic mycobacteria were found to assemble actin similarly to the LBP but those containing the pathogens M.tuberculosis or M.avium are known to be strongly inhibited in this process, in fusion with lysosomes and in acidification. Lipids were identified that, when added to infected cells could switch on, not only actin assembly by pathogenic mycobacterial phagosomes but also phagosomal maturation in general, leading to pathogen killing. A biocomputing approach, elementary mode analysis is being applied to identify the flux modes of metabolites through the network in the phagosomal membrane, leading to many predictions which could be experimentally verified. Our goal is to combine theory and experiments to provide a holistic description of how membrane signalling networks regulate two defined membrane functions, actin assembly and binding.

已经开发了两种体外试验来分析确定的细胞质膜表面，来自巨噬细胞的乳胶珠吞噬体（LBP）（A）如何从头组装肌动蛋白和（B）如何结合F-肌动蛋白。已经确定了调节组装过程的蛋白质和脂质的大型网络。发现非致病性分枝杆菌的吞噬体与LBP类似地组装肌动蛋白，但已知含有病原体结核分枝杆菌或鸟分枝杆菌的吞噬体在此过程中与溶酶体融合并酸化时受到强烈抑制。脂质被鉴定为，当被添加到感染的细胞中时，不仅可以通过致病性分枝杆菌吞噬体启动肌动蛋白组装，而且还可以通过一般的吞噬体成熟，导致病原体杀死。一种生物计算方法，基本模式分析被应用于确定通过网络的代谢物在吞噬体膜的通量模式，导致许多预测，可以通过实验验证。我们的目标是联合收割机的理论和实验提供一个全面的描述膜信号网络如何调节两个定义的膜功能，肌动蛋白组装和绑定。

项目成果

Dynamic life and death interactions between Mycobacterium smegmatis and J774 macrophages

• DOI: 10.1111/j.1462-5822.2005.00675.x
• 发表时间: 2006-06-01
• 期刊: CELLULAR MICROBIOLOGY
• 影响因子: 3.4
• 作者: Anes, E;Peyron, P;Griffiths, G
• 通讯作者: Griffiths, G

cAMP synthesis and degradation by phagosomes regulate actin assembly and fusion events: consequences for mycobacteria

• DOI: 10.1242/jcs.03091
• 发表时间: 2006-09-01
• 期刊: JOURNAL OF CELL SCIENCE
• 影响因子: 4
• 作者: Kalamidas, Stefanos A.;Kuehnel, Mark P.;Griffiths, Gareth
• 通讯作者: Griffiths, Gareth