Supplement to Chemical Biology Studies of the Dynamics and Inhibition of Peptidoglycan Biosynthesis

融资机会公告（FOA）编号：PA-20-272 最大化研究者研究奖R35 136365的设备申请（项目名称：化学 肽聚糖生物合成的动力学和抑制的生物学研究 项目摘要 这一设备要求正在作出显着提高我们的研究能力， 支持我们的R35 MIRA奖。我们资助的MIRA应用程序代表了两个融合 补充研究计划，概括地说，针对紧迫的公共卫生 抗生素耐药性带来的威胁。我们的MIRA奖旨在利用我们的发现， 荧光D-氨基酸（FDAA）提供了前所未有的，迄今为止无法获得的， 用于在真实的时间和现场中可视化细菌细胞壁肽聚糖（PG）动力学的工具 细菌细胞我们已经启动了额外的研究，以阐明细菌细胞分裂的细节， 枯草芽孢杆菌中的细胞分离，并且我们正在寻求开发新种类的荧光（即，“转─ on”）探针，其将使得能够实时和在活细菌中研究PG合成和动力学 细胞我们还将继续致力于环的合成和机理研究。 抑制PG生物合成的缩肽抗生素。此外，我们最近发现的数据 这表明环缩酚肽可能具有第二种作用机制， 脂质循环，PG生物合成途径中的一个重要活动。脂质再循环途径 仍然需要清楚地阐明，当与可能是固有的双模式活动相结合时， 这些正在研究的环缩酚肽，非常有前途的新途径，用于鉴定新的 抗菌目标和新的抗菌剂的开发很可能由此产生 研究工作。 这些领域的研究都需要常规使用高效液相色谱仪 分辨率能力与多种检测模式。我们目前的HPLC系统大约 20岁，不再受制造商的支持。此外，计算机系统 运行该仪器所需的时间已经过时，我们的大学不再支持。作为 结果，这种仪器有大量的“停机时间”，对研究进展产生不利影响 R35的奖励。在这个应用程序中，我们正在寻求资金购买安捷伦 1290 Infinity Flex系列HPLC/UHPLC仪器，能够进行常规纯度分析， 氨基酸、环肽等，以及UHPLC能力，能够分析更多 复杂样品，如PG片段。这种仪器将取代我们老化和无法修复的 系统，大大提高了我们的分析能力，并对我们的工作产生了深远的积极影响。 在R35奖励下的活动。

Funding Opportunity Announcement (FOA) Number: PA-20-272 Equipment request for Maximizing Investigators' Research Award R35 136365 (Project Title: Chemical Biology Studies of the Dynamics and Inhibition of Peptidoglycan Biosynthesis) PROJECT SUMMARY This equipment request is being made to significantly enhance our research capabilities in support of our R35 MIRA award. Our funded MIRA application represents the fusion of two complementary research programs that are, broadly described, directed at the urgent public health threat posed by antibiotic resistance. Our MIRA award seeks to capitalize on our discovery of fluorescent D-amino acids (FDAAs) that have provided unprecedented, and heretofore unavailable, tools for the visualization of bacterial cell wall peptidoglycan (PG) dynamics in real time and in live bacterial cells. We have initiated additional studies to elucidate the details of bacterial cell division and cell separation in Bacillus subtilis, and we are seeking to develop new classes of fluorogenic (i.e., “turn- on”) probes that will enable the study of PG synthesis and dynamics in real-time and in live bacterial cells. We will also continue our effort directed at the synthesis and mechanistic study of cyclic depsipeptide antibiotics that inhibit PG biosynthesis. Furthermore, we have recently uncovered data that suggest the cyclic depsipeptides may have a second mechanism of action that involves inhibition of lipid recycling, an essential activity in the PG biosynthesis pathway. The lipid recycling pathway remains to be clearly elucidated and, when coupled with dual-mode activity that may be inherent to these cyclic depsipeptides under study, very promising new avenues for the identification of new antibacterial targets and development of new antibacterial agents are likely to emerge from this research effort. Research in each of these areas requires routine access to HPLC instrumentation with high resolution capabilities with multiple mods of detection. Our current HPLC system is approximately twenty years old and is no longer supported by the manufacturer. In addition, the computer system required to run the instrument has become obsolete and is no longer supported by our university. As a result, this instrument has a significant amount of ‘down time’ that adversely impacts research progress under the parent R35 award. In this application, we are seeking funds for the purchase of an Agilent 1290 Infinity Flex Series HPLC/UHPLC instrument that has the capability for routine purity analysis of amino acids, cyclic peptides, and the like as well as UHPLC capability to enable analysis of more complex samples, such as PG fragments. This instrument will replace our aging and unrepairable system, significantly enhance our analytical capabilities, and have a profoundly positive impact on our activities under the parent R35 award.