Structures of Mtb proteins conferring susceptibility to known Mtb inhibitors

描述(申请人提供)：该项目的总体目标是确定潜在的药物靶点和抑制结核分枝杆菌(Mtb)和其他病原菌生长的伴生抑制剂，以便为抗菌药物的发现奠定基础。我们将通过识别结核分枝杆菌全细胞生长抑制物靶向的蛋白质来实现这一点，这是最近对20多万个类药物小分子进行的高通量筛选发现的。作为抑制剂靶标的蛋白质将通过遗传和互补生化方法进行鉴定。将与蛋白质结构倡议大型中心合作，确定作为抑制剂靶标的Mtb蛋白质或这些蛋白质的同系物的结构。在有人类同源物的情况下，也将确定该结构或相关真核蛋白的结构。结构信息和抑制物身份将与相关蛋白质的注释一起使用，以建议每个目标蛋白质的分子功能。将进行这些分子功能的分析，以确认功能和抑制。一旦确定了目标，将进一步分析它们的抑制剂对其他病原菌和真核细胞系的活性。然后，将根据目标的新颖性、伴随生长抑制物的活性光谱和抑制物的效力来确定目标的优先顺序。在五年的资助期间，我们希望确认大约200个顶级全细胞活性分子的分子靶标，并获得这些靶标蛋白质或相同序列家族成员的结构。这些结构、抑制物-蛋白质对以及有关蛋白质的生化信息将为结核分枝杆菌药物开发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，并为通过治疗结核病来改善人类健康的努力提供一条前进的道路。 公共卫生相关性：这项提议与人类健康的相关性在于，它将产生支持结核病药物发现所需的独特结构和功能数据，结核病仍然是全球主要的死亡原因。

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.