MAD PHASING AND HIGH RESOLUTION ANALYSIS OF BACTERIAL MEMBRANE PROTEINS

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。本方案正在进行细菌膜蛋白的新型晶体结构溶液的研究。我们已经结晶了III型分泌器的几个组件，它横跨病原菌的内膜和外膜，并允许细菌毒力蛋白进入宿主细胞。这种分泌器在几种革兰氏阴性病原体中高度保守，包括沙门氏菌、肠出血性大肠杆菌、假单胞菌、博达氏菌和志贺氏菌。将寻求EscN(细菌内膜分泌机构的一种基本ATPase)、EscJ和ESCC的复合体(被认为在细菌外膜形成一个大的环状结构)和EspB(被认为在人类宿主膜上形成孔)的硒蛋氨酸相变。此外，还将对转位到宿主细胞中的关键毒力因子进行分期，然后插入宿主细胞膜以促进细菌黏附(TIR用于转位的内膜受体)。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The novel crystallographic structure solution of bacterial membrane proteins is being undertaken in this proposal. We have crystallized several components of the type III secretion apparatus which spans the inner and outer membrane of pathogenic bacteria, and allows for the passage of bacterial virulence proteins into the host cell. This secretion apparatus is highly conserved amongst several gram negative pathogens including Salmonella, Enterhemmorhaghic E. coli, Psedomonas, Bordatella and Shigella. Seleno-methionine phasing of EscN (an essential ATPase of the secretion apparatus at the bacterial inner membrane), a complex of EscJ and EscC (which are presumed to form a large ring like structure in the bacterial outer membrane), and EspB (presumed to form the pore in the human host membrane) will be pursued. Additionally, the phasing of a key virulence factor that is translocated into the host cell, then inserted into host cell membranes to facilitate bacterial adhesion (TIR for translocated intimin receptor) will also be pursued.