Pathogen-specific drug targets for weaponized bacteria

描述（由申请人提供）：对当前抗生素治疗具有抗性的工程化细菌生物武器的威胁为开发新型治疗化合物提供了强有力的授权。该提案提出了洛克菲勒大学三个互补小组（Vincent Fischetti博士，亚历山大Tomasz博士和Erec Stebbins博士）的共同努力，以确定A类病原体B的新型抑制剂。anthracis和Y.鼠疫以及B类食品和水安全威胁。第一个也是中心研究项目是基于观察到对噬菌体裂解酶如PlyG对B的抗性。炭疽杆菌是极其罕见的事件，由此我们假设这些酶的结合结构域靶向细菌细胞壁中的关键组分以确保噬菌体释放（在通过进化的意义上，噬菌体已经进行了高通量测定以鉴定该组分）。因此，合成这种壁组分的途径代表了抗生素开发的目标。因此，核心项目将侧重于裂解酶结合底物的鉴定、分离和表征。一旦确定，将确定其合成途径。催化该途径的酶将被分离、纯化和结晶，并鉴定抑制剂。Alex Tomasz的细胞壁结构经验为核心项目提供了信息，但也将其扩展到寻找针对B的其他新靶点。炭疽病的细胞壁的高分辨率分析和条件致死突变体的分离-使用水手型元件的体内转座。细胞壁在他的项目中占据中心地位，因为（i）PlyG的靶点在细胞壁中;（ii）细胞壁在B引起的毁灭性败血性休克中起着重要作用。炭疽菌感染和（iii）B. B.炭疽菌含有壁相关决定子，这些决定子是其他细菌中的毒力因子。Erec Stebbins将提供蛋白质结构和结晶方面的专业知识，沿着这些结构的小分子抑制剂的鉴定。他的项目扩大了新疗法的研究范围，将Y。鼠疫和B类病原体通过计算药物发现中的翻译组件耦合到结构确定。总体而言，这些努力针对关键生物战病原体的核心毒力因子。

DESCRIPTION (provided by applicant): The threat of engineered bacterial bioweapons that are resistant to current antibiotic treatments provides a powerful mandate to develop novel classes of therapeutic compounds. This proposal presents a concerted effort of three complementary groups at the Rockefeller University (Dr. Vincent Fischetti, Dr. Alexander Tomasz, and Dr. Erec Stebbins) to identify novel inhibitors of the Category A pathogens, B. anthracis and Y. pestis, as well as Category B food and water safety threats. The first and central research project is based on the observation that resistance to bacteriophage lytic enzymes, such as PlyG for B. anthracis, is an extremely rare event, from which we hypothesize that the binding domain of these enzymes targets a critical component in the bacterial cell wall to assure phage release (in a sense through evolution, the phage have performed the high throughput assay to identify this component). Thus, the pathway to the synthesis of this wall component represents a target for antibiotic development. The core project will therefore focus on the identification, isolation and characterization of the lytic enzyme binding substrate. Once identified, the pathway towards its synthesis will be determined. Those enzymes catalyzing the pathway will be isolated, purified and crystallized, and inhibitors identified. The cell wall structural experience of Alex Tomasz, feeds into the core project but also broadens it to a search for additional novel targets against B. anthracis by high resolution analysis of the cell walls and by the isolation of conditional lethal mutants - using in vivo transposition of a mariner type element. Cell walls occupy center stage in his project since (i) the target of PlyG is in the wall; (ii) cell walls play an important role in the devastating septic shock caused by B. anthracis infections and (iii) the genome of B. anthracis contains wall-related determinants that are virulence factors in other bacteria. Erec Stebbins will provide the expertise in protein structure and crystallization along with the identification of small molecule inhibitors for these structures. His project expands the novel therapeutics search to include Y. pestis and Category B pathogens through a translational component in computation drug discovery coupled to structural determination. In total, these efforts target central virulence factors in critical biowarfare pathogens.