Struktur und Interaktion von Proteinen aus dem Reggie-like Operon von Prokaryoten / Structure and Interaction of Proteins of the Reggie-like Operon in Prokaryotes

由于这些膜相关蛋白及其构建体的不利表达和体外行为，对真核reggie蛋白的结构研究迄今为止极其困难。因此，本项目的重点是原核蛋白质编码的雷吉样操纵子，其特征在于核磁共振光谱。雷吉样蛋白（RLP）在许多细菌中发现。最有趣的是，这些RLP几乎总是伴随着属于NfeD家族的另一种跨膜蛋白。在一些情况下，另外的雷吉相关蛋白（RAP）在雷吉样操纵子内编码，并因此与RLP共表达和共调节。在B。在枯草芽孢杆菌中，雷吉样操纵子受σ W因子调节，导致在细胞应激条件下（如渗透压休克或存在细胞毒性损害抗生素）表达增强。看来，RLP和它们相关的伴侣蛋白共同作用于这些应激条件。原核RAP和RLP的高分辨率结构以及它们的分子相互作用的表征，这是本提案的主要目标，将是理解它们的功能以及如何与它们的真核同源物相关的关键。

Structural studies on eukaryotic reggie proteins have so far been extremely difficult due to the adverse expression and in vitro behavior of these membrane-associated proteins as well as constructs thereof. The present project focuses therefore on prokaryotic proteins encoded in the reggie-like operons which will be characterized by NMR spectroscopy. Reggie-like proteins (RLPs) are found in many bacteria. Most interestingly, these RLPs are almost always accompanied by another transmembrane protein belonging to the NfeD family. In some cases further reggie-associated proteins (RAPs) are encoded within the reggie-like operon and thus co-expressed and co-regulated with the RLPs. In B. subtilis, the reggie-like operon is regulated by the σW-factor leading to enhanced expression under conditions of cellular stress, like osmotic shock or presence of cell envelope-compromising antibiotics. It appears that RLPs and their associated partner proteins altogether act in response to these stress conditions. High-resolution structures of prokaryotic RAPs and RLPs as well as characterization of their molecular interactions, which are the major objectives of this proposal, will be key to understanding their function and how this is related to their eukaryotic homologs.