Project 3 - The Critical Role of Membrane Transport

项目摘要-项目3-以膜转运蛋白的关键作用为目标 结核分枝杆菌是一种细胞内病原体，因此被宿主细胞包围。因为这种身临其境的 在环境中，结核分枝杆菌膜蛋白构成了病原体和细菌之间的直接功能界面。 它的主人。它们感知环境并控制营养物质和其他分子(包括许多 毒品)进入和离开有机体。因此，它们为小分子疗法提供了一条“大门”。 然而，到目前为止，只有两种Mtb整膜蛋白的高分辨结构是由于 部分原因是膜蛋白的表达、纯化和结晶困难，以及缺乏 将重点放在Mtb上。我们开发了一种通用的方法来确定完整的膜蛋白的优先顺序 在结核分枝杆菌中被确定为有效的药物靶点、基本基因或关键毒力因子。我们的优先事项继续 与财团成员合作，使用初步药物靶向和 毒力筛选数据。基于我们作为国家中心发展起来的专业知识，我们还 开发了表达、纯化和结晶用于结构的完整膜蛋白的可靠方法 用X射线结晶学和电子冷冻显微镜进行测定。

Project Summary – Project 3 – Targeting the Critical Role of Membrane Transporters Mtb is an intracellular pathogen and thus is surrounded by a host cell. Because of this immersive environment, Mtb membrane proteins constitute a direct functional interface between the pathogen and its host. They sense the environment and control entry of nutrients and other molecules (including many drugs) into, and exit from the organism. Thus, they offer a ‘gateway’ for small molecule therapeutics. However, to date, there are only two high-resolution structures of Mtb integral membrane proteins due in part to the difficulty of expression, purification and crystallization of membrane proteins, and to the lack of focus onto Mtb. We have developed a general method to prioritize integral membrane proteins identified as viable drug targets, essential genes, or critical virulence factors in Mtb. Our priorities continue to be ranked in collaboration with members of the consortium, using preliminary drug targeting and virulence screening data. Building on our expertise developed as a national center, we have also developed robust methods to express, purify and crystallize integral membrane proteins for structure determination by X-ray crystallography, and by electron cryo-microscopy.