Structures of Mtb proteins conferring susceptibility to known Mtb inhibitors

描述（由申请人提供）：该项目的总体目标是确定抑制结核分枝杆菌（Mtb）和其他致病菌生长的潜在药物靶点和伴随抑制剂，以便为抗菌药物的发现奠定基础。我们将通过鉴定被结核分枝杆菌全细胞生长抑制剂靶向的蛋白质来实现这一目标，这些抑制剂是最近对超过20万种药物样小分子进行的高通量筛选发现的。作为抑制剂靶点的蛋白质将通过遗传和互补的生化方法来鉴定。与蛋白质结构倡议大规模中心合作，将确定作为抑制剂靶点的结核分枝杆菌蛋白或这些蛋白的同源物的结构。在人类同源物可用的情况下，该结构或相关真核蛋白的结构也将被确定。结构信息和抑制剂身份将与相关蛋白的注释一起使用，以提示每个靶蛋白的分子功能。将对这些分子功能进行测定，以确认其功能和抑制作用。一旦目标被确定，它们的抑制剂将进一步分析它们对其他致病菌和真核细胞系的活性。然后根据它们的新颖性、伴随生长抑制剂的活性谱和抑制剂的效力对靶标进行优先排序。在五年的资助期间，我们预计将确认大约200个顶级全细胞活性分子的分子靶标，并获得这些靶蛋白或同一序列家族成员的结构。这些结构、抑制剂-蛋白对以及蛋白质的生化信息将为结核分枝杆菌药物的发现提供丰富的先导化合物和生物化学资源，为了解结核分枝杆菌生物学奠定基础，并为通过治愈结核病改善人类健康的努力提供前进的道路。

DESCRIPTION (provided by applicant): The overall goal of this project is to identify potential drug targets and companion inhibitors that inhibit growth of Mycobacterium tuberculosis (Mtb), and other pathogenic bacteria in order to develop a foundation for antimicrobial drug discovery. We will accomplish this by identifying proteins that are targeted by whole cell growth inhibitors of Mtb discovered by a recently conducted high-throughput screen of over 200,000 drug-like small molecules. Proteins that are targets of inhibitors will be identified by genetic and complementary biochemical approaches. Structures will be determined, in collaboration with the Protein Structure Initiative Large-Scale Centers, of either the Mtb proteins that are targets of inhibitors or homologs of these proteins. In cases where a human homolog is available, that structure or that of a related eukaryotic protein will be determined as well. The structural information and inhibitor identities will be used along with annotations of related proteins to suggest molecular functions for each target protein. Assays for these molecular functions will be carried out to confirm both function and inhibition. Once targets are identified their inhibitors will additionally be analyzed for their activity against other pathogenic bacteria and eukaryotic cell lines. The targets will then be prioritized based on their novelty, the spectrum of activity of the companion growth inhibitor and the potency of the inhibitor. During the five years of the grant we expect to confirm the molecular targets for about 200 of the top whole cell active molecules and obtain the structures of these target proteins or members of the same sequence family. These structures, inhibitor-protein pairs, and biochemical information about the proteins will provide a rich resource of lead compounds and biochemistry for Mtb drug discovery, a foundation for understanding Mtb biology, and a path forward in the effort to improve human health by curing tuberculosis. PUBLIC HEALTH RELEVANCE: The relevance of this proposal to human health is that it will generate unique structural and functional data needed to support drug discovery for tuberculosis, which remains a leading cause of death worldwide.