Signalling-Pathways Regulating the Dynamic Localization of Z-disc-Associated Proteins

在过去的几年里，与肌原纤维z盘相关的蛋白质数量出乎意料地高。此外，已经发现编码许多Z-disc蛋白的基因突变可引起神经肌肉和/或心脏疾病。然而，对于调节特定蛋白-蛋白相互作用和z盘动态状态的信号通路知之甚少。现有的蛋白质相互作用图谱来源于生化工作，为使用最先进的显微镜方法系统分析动态蛋白质定位和相互作用提供了良好的基础。因此，我们将在光漂白后应用荧光显微镜（FRAP）和双分子荧光互补（BiFC）测量，并结合信号通路的直接干扰。该项目的见解对于蛋白质相互作用数据的解释（P2, P3, P4, P10），蛋白质复合物的结构确定，分析和验证（P5, P6）， z -盘超微结构分析和解释（P7）以及z -盘蛋白复合物和肌瘤动力学的生物物理表征（P8, P9）具有直接意义。

The last few years have witnessed the identification of an unexpectedly high number of proteins associated with the myofibrillar Z-disc. In addition it became evident that mutations in the genes encoding many Z-disc proteins have been found to cause neuromuscular and/or cardiac disease. However, very little is known about the signalling pathways that regulate specific protein-protein interactions and the dynamic state of the Z-disc. The existing map of protein interactions derived from biochemical work provides an excellent basis for a systematic analysis of dynamic protein localization and interactions by using state-of-the-art microscopy methods. We will therefore apply fluorescence microscopy after photobleaching (FRAP) and bimolecular fluorescence complementation (BiFC) measurements in combination with direct interference of signalling pathways. Insights from this project are of immediate importance for the interpretation of protein interaction data (P2, P3, P4, P10), structure determination, analysis and validation of protein complexes (P5, P6), Z-disc ultrastructure analysis and interpretation (P7) and biophysical characterization of Z-disc protein complexes and sarcomer dynamics (P8, P9).