STRUCTURAL STUDIES ON THE ROLE OF ZINC IN BACTERIAL PROLIFERATION

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 我们的INBRE支持的研究涉及一系列参与细菌增殖的金属酶中锌位置的结构特征。正在研究的系统包括一系列广泛的金属-β-内酰胺酶，它们对青霉素类抗生素具有耐药性。与相关的丝氨酸活性内酰胺酶不同，金属β-内酰胺酶没有已知的抑制剂，其中许多是以质粒编码的，允许在细菌物种之间轻松转移。还在研究的还有一系列AHL-内酰胺酶，它们作用于革兰氏阳性细菌的群体感觉通路，使细菌菌落表现出群体行为。我们利用X射线吸收光谱直接询问冷冻溶液中的金属位置，而不需要结晶材料。我们的方法提供了天然金属离子的酶休眠状态和产物络合物，并通过快速冷冻淬火技术，关键的反应中间体

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Our INBRE supported research involves the structural characterization of the zinc sites in a series of metalloenzymes involved in bacterial proliferation. The systems under study include a broad array of metallo-b-lactamases, which confer resistance to penicillin-type antibiotics. Unlike the related serine-active lactamases, the metallo-b-lactamases have no known inhibitors and many of them are plasmid encoded, allowing for facile transfer between bacterial species. Also under study are a series of AHL-lactonases which act in the quorum-sensin pathways of gram-positive bacteria, allowing bacterial colonies to display group behavior. We utilize X-ray absorption spectroscopy to directly interrogate the metal sites in frozen solution, without the need for crystalline material. Our methodology provides access to the native metal ion in enzyme resting states and product complexes, and through rapid freeze quench techniques, key reaction intermediates