Mapping the structural basis for mechanistic diversity in metalloenzyme superfamilies

项目摘要 这种补充剂的父母拨款旨在发现与dna生物合成有关的新的酶辅助因子。 在病原体中。首要目标是为抑制引起传染病的细菌中的酶提供新的途径。 疾病。在这里，我们将研究一种必需的酪氨酸衍生的翻译后组装的机制 在微生物核糖核苷酸还原酶的一个亚类中发现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确定可以作为目标的辅因子组装的各个方面 通过治疗干预进行干扰。在第二个目标中，我们将描述这种酶所使用的辅因子。 原生寄主生物体中的亚纲。作为一个整体，该项目将为理解这种酶如何 子类在细胞内使用，最终允许我们测试辅因子组装中断的影响 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 在化学和微生物学之间的工作中的补充支持。协作型 指导团队具备非天然氨基酸方法学、结构和生化方面的专业知识 蛋白质的特性，以及病原微生物和共生微生物的培养。

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.