Natural compounds target the M23B zinc metallopeptidase Mpg to modulate Neisseria gonorrhoeae Type IV pilus expression

Neisseria gonorrhoeae uses the Type IV pilus (T4p) to colonize several sites within humans by adhering to host cells and tissues. Previously, we identified a periplasmic M23B zinc metallopeptidase, Mpg, that is necessary to protect from oxidative and nonoxidative killing and these phenotypes are mediated by Mpg activities on T4p expression. Here, we use a high-throughput, target-based screening approach to identify novel inhibitors of Mpg's enzymatic activity. We identified two natural compounds, punicalagin and chebulinic acid, which inhibit the peptidoglycan-hydrolyzing activity of Mpg in a dose-dependent manner. Moreover, treatment of N. gonorrhoeae with these compounds leads to a concomitant decrease in the number of T4p, similar to an mpg mutant. However, these compounds are not toxic to N. gonorrhoeae. These compounds exhibit activity against Mpg orthologs from other Bacterial species. Notably, these natural compounds inhibit N. gonorrhoeae colonization and survival in Cell Culture models of Infection. This work provides the characterization of two natural compounds with activity against N. gonorrhoeae T4p through the Mpg M23B class zinc metallopeptidase.

Importance: Neisseria gonorrhoeae is a global health burden with high transmission rates and multidrug resistance. N. gonorrhoeae encodes a Type IV pilus (T4p), which is a major colonization and virulence factor. The importance of the T4p in multiple stages of Infection makes it an attractive drug target. Previously, we identified an M23B zinc metallopeptidase, Mpg, important for T4p production and T4p-mediated resistance to neutrophil killing. In this study, we identified two natural compounds, punicalagin and chebulinic acid, as novel inhibitors of Mpg's enzymatic activity that thus inhibit T4p expression. These findings identify two potential anti-colonization and anti-virulence compounds and provide a framework to target T4p components for future screens, poising the field to potentially discover additional compounds to combat N. gonorrhoeae Infection.

Neisseria gonorrhoeae; metallopeptidase; natural compounds; pilus; small molecule screen.