Signalling networks in latex bead- and mycobacterial-phagosomes: Analysis of dynamics and modulation of actin-membrane assembly

乳胶珠和分枝杆菌吞噬体中的信号网络：肌动蛋白膜组装的动力学和调节分析

• 批准号: 5455586
• 负责人: Privatdozent Dr. Sergei A. Kuznetsov
• 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: 2005
• 资助国家: 德国
• 起止时间: 2004-12-31 至 2006-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/5455586?language=en
• 关键词: Signalling; networks; latex; bead; mycobacterial

In vitro assays analyse how a defined cytoplasmic membrane surface, latex bead phagosomes (LBP) from macrophages (A) assembles actin de novo and (B) binds to F-actin. In the first period we found (i) a large network of proteins and lipids that switch on actin assembly and maturation also in pathogenic mycobacterial phagosomes; (ii) a biocomputing approach identified flux modes of metabolites through the membrane, many predictions could be experimentally verified, including the synthesis of ATP; (iii) a time-step simulation to predict effects of lipid combinations on phagosomal actin under different conditions; (iv) involvement of prostaglandins and leukotrienes in actin nucleation and (v) regulation of phagosome binding to actin by arachidonate is regulated in opposite manner to actin nucleation. New work will focus on these new aspects to combine theory and experiment to provide a holistic description how these membrane signalling networks regulate actin assembly and binding: (i) kinetics and concentrations in the process of actin nucleation for key compounds; (ii) involvement of prostaglandins and other arachidonic metabolites in actin nucleation; (iii) the signalling network regulating phagosome binding to actin.

体外测定分析确定的细胞质膜表面、巨噬细胞的乳胶珠吞噬体 (LBP) (A) 如何从头组装肌动蛋白以及 (B) 与 F-肌动蛋白结合。在第一阶段，我们发现（i）在致病性分枝杆菌吞噬体中也存在一个巨大的蛋白质和脂质网络，可以开启肌动蛋白的组装和成熟； (ii) 生物计算方法确定了代谢物通过膜的通量模式，许多预测可以通过实验验证，包括 ATP 的合成； (iii) 时间步长模拟，以预测不同条件下脂质组合对吞噬体肌动蛋白的影响； (iv)前列腺素和白三烯参与肌动蛋白成核，以及(v)花生四烯酸对与肌动蛋白结合的吞噬体的调节以与肌动蛋白成核相反的方式调节。新的工作将集中在这些新方面，将理论和实验结合起来，提供这些膜信号网络如何调节肌动蛋白组装和结合的整体描述：（i）关键化合物肌动蛋白成核过程中的动力学和浓度； (ii) 前列腺素和其他花生四烯酸代谢物参与肌动蛋白成核； (iii) 调节吞噬体与肌动蛋白结合的信号网络。