MAD PHASING AND HIGH RESOLUTION ANALYSIS OF BACTERIAL MEMBRANE PROTEINS

细菌膜蛋白的疯狂定相和高分辨率分析

• 批准号: 7370382
• 负责人: NATALIE CJ STRYNADKA
• 金额: $ 0.58万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-03-01 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7370382
• 关键词: MAD; PHASING; RESOLUTION; ANALYSIS; BACTERIAL

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.

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。该方案正在进行细菌膜蛋白的新晶体学结构解决方案。我们已经结晶了III型分泌装置的几个组件，其跨越病原菌的内膜和外膜，并允许细菌毒力蛋白进入宿主细胞。这种分泌装置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。大肠杆菌、假单胞菌、博德特氏菌和志贺氏菌。将追踪EscN（细菌内膜处分泌装置的必需ATP酶）、EscJ和EscC的复合物（推测其在细菌外膜中形成大的环状结构）和EspB（推测其在人类宿主膜中形成孔）的硒-甲硫氨酸定相。此外，还将对易位到宿主细胞中，然后插入宿主细胞膜以促进细菌粘附的关键毒力因子进行定相（易位的内膜受体的TIR）。