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结构的不良表达和体外行为，真核reggie蛋白的结构研究迄今为止非常困难。因此，本项目的重点是编码雷吉样操纵子的原核蛋白，这些蛋白将用核磁共振波谱法进行表征。雷吉样蛋白（rlp）存在于许多细菌中。最有趣的是，这些rlp几乎总是伴随着另一种属于NfeD家族的跨膜蛋白。在某些情况下，进一步的reggie-associated protein （RAPs）被编码在reggie-样操纵子内，因此与rlp共表达和共调节。在枯草芽孢杆菌中，reggilike操纵子受σ w因子的调控，导致在渗透休克或破坏包膜的抗生素存在等细胞应激条件下表达增强。似乎rlp及其相关的伴侣蛋白共同对这些应激条件作出反应。原核RAPs和rlp的高分辨率结构及其分子相互作用的表征是本研究的主要目标，这将是了解它们的功能及其与真核同源物之间关系的关键。

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.