Co-translational protein insertion into the bacterial plasma membrane (P16)

整合膜蛋白以共翻译的方式通过SecYEG易位子（质膜中的蛋白传导通道）的侧门插入细菌质膜中。辅助蛋白YidC、SecDF和YajC与核心易位子SecYEG一起形成全易位子。本项目的长期目标是了解易位子作为内膜蛋白生物合成的细胞门的功能。我们将使用主要基于荧光的方法，包括PET、FCS、FRET和单分子FRET，在真实的时间内跟踪共翻译膜插入。为了研究YidC如何被纳入，我们将研究SecYEG在YidC的N-末端区域的膜插入和YidC的C-末端结构域从核糖体到SecB/SecA的移交中的作用。不同拓扑结构的蛋白质的共翻译折叠和膜插入将使用LepB和EmrD作为不同拓扑结构排列的示例进行监测。然后，我们将研究YidC和SecYEG如何合作插入膜蛋白，特别是多位膜蛋白。这些实验将提供一个动态和功能的细胞门守卫插入，组装和拓扑结构的内膜蛋白质的洞察。

Integral membrane proteins insert into the bacterial plasma membrane in a co-translational fashion through the lateral gate of the SecYEG translocon, a protein-conducting channel in the plasma membrane. Together with the core translocon SecYEG the accessory proteins YidC, SecDF and YajC form the holotranslocon. The long-term aim of this project is to understand the function of the translocon as a cellular gate for the biogenesis of inner-membrane proteins. We will follow co-translational membrane insertion in real time using mainly fluorescence-based methods, including PET, FCS, FRET, and single-molecule FRET. To study how YidC is incorporated we will examine the role of SecYEG in the membrane insertion of the N-terminal region of YidC and the handover of the C-terminal domain of YidC from the ribosome to SecB/SecA. Co-translational folding and membrane insertion of proteins of different topology will be monitored using LepB and EmrD as examples for different topological arrangements. We will then examine how YidC and SecYEG cooperate in the insertion of membrane proteins, in particular of polytopic membrane proteins. These experiments will provide an insight into dynamics and function of a cellular gate guarding insertion, assembly and topology of inner-membrane proteins.