RUI: Chemical probes for studying peptidoglycan metabolism

点击翻译按钮获取中文摘要

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.