Analyse der Sekundärmodifikationen von Komponenten der Phytochrom-Signaltransduktion in Arabidopsis thaliana

迄今为止，复杂信号转导网络的绘制主要依赖于突变体的分析、体外和体内相互作用的研究以及单个组分的生化表征。现代质谱法允许鉴定单一蛋白质和复杂混合物中肽的二级修饰的表征。这允许对已知信号组件的状态进行时间解析分析。光敏色素A （phyA）的信号通路可以由远红光特异性触发，因此独特地允许观察具有许多已知成分的单个信号链，将使用所提出的技术作为目标。该项目以及与Baginsky （Halle）的计划提案旨在检测复杂蛋白质组样品中关键phyA信号成分的空间和时间二次修饰，从而借助SRM（选择反应监测）质谱（LC-ESI-MS/MS）阐明关键成分的相互作用。

Mapping of complex signal transduction networks has so far mainly relied on the analysis of mutants, studies of interactions in vitro and in vivo and on the biochemical characterisation of single components. Modern mass spectrometry allows the identification of single proteins and the characterisation of secondary modifications of peptides in complex mixtures. This allows a timeresolved analysis of the status of known signalling components. The signalling pathway of phytochrome A (phyA) which can specifically be triggered by far-red light thus uniquely allowing the observation of a single signalling chain with many known components will be targeted using the proposed technique. The project and a planned proposal with Baginsky (Halle) aim to detect spacial and temporal secondary modifications of key phyA signalling components in complex proteome samples and hence to elucidate the interplay of the key components with the help of SRM (selected reaction monitoring) mass spectrometry (LC-ESI-MS/MS).