RUI: Chemical probes for studying peptidoglycan metabolism

有了这个奖项，化学部的生命过程化学计划正在资助克里斯托弗·W。来自布莱恩特大学的里德开发了化学工具来研究细菌的细胞壁。虽然我们对新细胞壁材料是如何构建的知识相对比较成熟，但我们对拆卸过程以及细胞壁拆卸与组装的协调的理解却不太为人所知。小分子抑制剂通常用于研究生物过程。降解细胞壁多糖骨架的酶的小分子抑制剂为研究细胞壁组装和拆卸之间的相互作用提供了新的途径。实验程序有助于表征这些酶的化学抑制和遗传失活之间的分子差异。该项目允许本科生和学士后研究员在化学和生物学的界面上获得多学科培训。研究细胞壁降解的新分子和方法将提供给科学界，并可能有助于提高我们对细胞壁组装和拆卸的理解。该项目被纳入一个推广计划，向高中生介绍化学生物学。 里德教授致力于促进对话和参与科学院外，并参加了几个活动在当地学校（K-12），无论是正式（国家科学博览会，RI-NSF-EPSCoR），和非正式（在小学志愿服务）。 他计划通过提供暑期研究奖学金和指导来自经济困难学区的代表性不足的二年级和初中学生来扩大他与罗得岛高中的联系。这个研究项目对降解细菌细胞壁多糖骨架的酶的化学和遗传失活之间的差异进行定量表征。该项目结合了遗传学、生理学和生物物理学方法，以验证模式生物枯草芽孢杆菌中一类称为N-乙酰氨基葡萄糖苷酶（GlcNAcases）的自溶素的小分子抑制剂。该研究项目将使用基于发现的方法（定量PCR，定点诱变）和生物物理表征（差示扫描荧光法，内在蛋白质荧光）来研究这些酶化学灭活后观察到的细胞和表型变化。 这项研究的信息可能提供新的方法和试剂，可用于更好地了解一类对细菌细胞壁生长很重要的酶的结构和功能。验证这些抑制剂作为探针研究细胞壁作用GlcNAcases可以提供深入了解细菌自溶素的遗传和化学灭活之间的生理差异及其对细胞壁metabolic.This奖项反映了NSF的法定使命，并已被认为是值得通过使用基金会的智力价值和更广泛的影响审查标准进行评估的支持。

With this award, the Chemistry of Life Processes Program in the Chemistry Division is funding Dr. Christopher W. Reid from Bryant University to develop chemical tools to study the cell wall of bacteria. While our knowledge of how new cell wall material is built is relatively well established, our understanding of the disassembly process and the coordination of cell wall disassembly with assembly is less well known. Small molecule inhibitors are often used to study biological processes. Small molecule inhibitors of enzymes that degrade the polysaccharide backbone of the cell wall provide a new approach to study the interaction between the assembly and disassembly of the cell wall. The experimental procedures help to characterize the molecular differences between chemical inhibition and genetic inactivation of these enzymes. This project allows undergraduate students and post-baccalaureate fellows to acquire multi-disciplinary training at the interface of chemistry and biology. The new molecules and methods for studying the degradation of the cell wall will be made available to the scientific community, and may help improve our understanding of cell wall assembly ands disassembly. This project is integrated into an outreach program to introduce high school students to chemical biology. Professor Reid is committed to fostering dialog and engagement with science outside of the academy, and has participated in several activities at local area schools (K-12), both formally (state science fair, RI-NSF-EPSCoR), and informally (volunteering at elementary schools). He plans to expand his association with high schools in Rhode Island by providing summer research fellowships and mentoring to under-represented sophomore and junior high school students from economically challenged school districts.This research project undertakes a quantitative characterization of the differences between chemical and genetic inactivation of enzymes that degrade the polysaccharide backbone of the bacterial cell wall. This project combines genetic, physiological, and biophysical approaches to validate a small molecule inhibitor of a class of autolysins known as N-acetylglucosaminidases (GlcNAcases) in the model organism Bacillus subtilis. This research project will use discovery-based approaches (quantitative PCR, site-directed mutagenesis), and biophysical characterization (differential scanning fluorimetry, intrinsic protein fluorescence) to investigate the cellular and phenotypic changes observed upon chemical inactivation of these enzymes. Information from this study may provide new methods and reagents that can be used to better understand the structure and function of a class of enzymes important to bacterial cell wall growth. The validation of these inhibitors as probes to study cell wall acting GlcNAcases may provide insight into the physiological differences between genetic and chemical inactivation of bacterial autolysins and the impact it has on cell wall metabolism.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.