Enzymes exist in crowded cellular environments. However, conventional laboratory studies are typically conducted in dilute conditions that do not account for the molecular crowding effects on enzyme structure, dynamics, and function. Our research focuses on investigating the crowding effects on the structure and stability of a multidomain enzyme, prolyl-tRNA synthetase (ProRS) from Escherichia coli. ProRS catalyzes the ligation of proline to tRNAPro, a crucial step in protein biosynthesis. We are using Atomic Force Microscopy (AFM), a type of scanning probe microscopy that produces high-resolution topographic images at the nanometer scale, to analyze protein structural changes. AFM images of ProRS in crowded environments will be presented. The results of this study are significant for rational drug design.
