Determining The Crystal Structure of The Staphylococcus Aureus MurG Enzyme

Patrick Wood head shot

Name: Patrick Thomas Wood
Major: Biochemistry and Molecular Biology
Minor: Statistical & Data Sciences
Advisors: Sara E.S. Martin, Annastassia Gallo

Research Supported by the Copeland Fund

Antibiotic-resistant bacteria are one of the biggest problems the human and veterinary healthcare industries face. As resistance to common-use antibiotics grows, so does the need to discover novel targets and develop new drugs against them. A potential new target is the MurG enzyme, a glycotransferase that synthesizes the bacterial cell wall. I propose that an inhibitor of MurG, the rate-limiting enzyme in the peptidoglycan biosynthesis pathway, could work as a novel antibiotic. Structural data is needed to develop a novel inhibitor. While structural data for two MurG enzymes have been solved using x-ray crystallography, no structural data exist for MurG in Staphylococcus aureus. Sequence similarity between MurG isoforms is generally low, and S. aureus MurG is one of 20% of MurG isoforms that have a phenylalanine variation in the active site, a variant yet to be crystallized. This work details an optimized expression and purification method for S. aureus MurG and describes a range of reproducible crystallization conditions. Additionally, this work attempts to further optimize human involvement in the crystal screening process through the use of an automated pipetting robot. Further experimentation is required to obtain large enough high-quality crystals suitable for x-ray diffraction. Structural data gained from x-ray diffraction would allow for the future development of specific inhibitors based on a quinolinone-6-sulfonamide (Q6S) moiety. Previous literature has shown that the Q6S moiety can mimic the binding of UDP-GlcNAc in other glycotransferases.

Posted in Comments Enabled, Independent Study, Symposium 2023 on April 14, 2023.

One response to “Determining The Crystal Structure of The Staphylococcus Aureus MurG Enzyme”

  1. Eli says:

    good stuff kid