Mapping the structural basis for mechanistic diversity in metalloenzyme superfamilies

项目摘要 该补助金旨在发现参与DNA生物合成的新型酶辅助因子 在病原体中。总体目标是使新的途径，酶抑制细菌，造成传染性 疾病在这里，我们将研究一个重要的酪氨酸衍生的装配后的机制， 在微生物核糖核苷酸还原酶亚类中发现的一种经修饰的自由基辅因子，通过掺入非- 天然氨基酸这种方法将使我们能够识别辅因子组装的各个方面， 治疗干预的破坏。在第二个目标中，我们将描述这种酶所使用的辅因子的特征 原生宿主有机体中的一个亚类。整个项目将为理解这种酶如何 子类在细胞内使用，最终使我们能够测试在细胞中破坏辅因子组装的影响。 相关的生物学背景。该项目为博士生提供了一个极好的培训机会。候选 在化学和微生物学之间的界面工作的补充支持。一个协作 指导团队具备非天然氨基酸方法学、结构和生物化学方面的专业知识 蛋白质的表征，以及病原微生物和真菌的培养。

Project Summary The parent grant for this supplement aims to discover novel enzymatic cofactors involved in DNA biosynthesis in pathogens. The overarching goal is to enable new routes to enzyme inhibition in bacteria that cause infectious disease. Here we will investigate the mechanism of assembly of an essential tyrosine-derived post-translationally modified radical cofactor found in a subclass of microbial ribonucleotide reductases through incorporation of non- native amino acids. This approach will allow us to identify aspects of cofactor assembly that could be targeted for disruption by therapeutic interventions. In a second aim, we will characterize the cofactor used by this enzyme subclass in a native host organism. The project as a whole will set the stage for understanding how this enzyme subclass is used inside the cell, ultimately allow us to test the impact of disruption of cofactor assembly in a relevant biological context. The project provides an excellent training opportunity for the Ph.D. candidate supported by the supplement in work at the interface between chemistry and microbiology. A collaborative mentoring team is in place with expertise in non-native amino acid methodology, structural and biochemical characterization of proteins, and cultivation of pathogenic and commensal microbes.