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

已经开发了两种体外试验来分析来自巨噬细胞的特定细胞质膜表面，乳胶头吞噬体（LBP） (a)如何重新组装肌动蛋白和(B)与f -肌动蛋白结合。一个由蛋白质和脂质组成的大网络已经被确定，它调节着组装过程。发现非致病性分枝杆菌的吞噬体与LBP相似地组装肌动蛋白，但已知含有结核分枝杆菌或鸟分枝杆菌病原体的吞噬体在与溶酶体融合和酸化过程中强烈抑制这一过程。研究发现，当脂质被添加到感染细胞中时，不仅可以激活致病性分枝杆菌吞噬体的肌动蛋白组装，还可以促进吞噬体的成熟，从而杀死病原体。一种生物计算方法，基本模式分析被应用于识别代谢物通过吞噬体膜网络的通量模式，导致许多可以实验验证的预测。我们的目标是将理论和实验结合起来，全面描述膜信号网络如何调节两种定义的膜功能，肌动蛋白组装和结合。

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.