We have developed a method to look for molecules that could be used as antibiotics, antivirulence therapeutics or anticoagulants using E. coli cells. After several rounds of screening of several synthetic small molecule libraries we have found few candidate inhibitors of the DsbB and VKOR enzymes of pathogenic bacteria. We will continue searching for more effective inhibitors of DsbB and VKOR as well as to ellucidate their mechanisms of inhibition. We are interested in expanding our inhibitor search using natural products libraries and molecules derived from the microbiome given their chemical diversity.
Here are the steps we took in our previous high throughput screenings (HTS) to find inhibitors of the disulfide bond forming enzymes of Pseudomonas aeruginosa and Mycobacterium tuberculosis and the steps to characterize them. We will apply similar strategies using more diverse libraries.
The goal of this project is to enable the systematic identification of substrates of the disulfide bond formation pathway of two human pathogens, Pseudomonas aeruginosa and Mycobacterium tuberculosis. We will develop a substrate profiling method using split fluorescent proteins to identify the proteins that interact with DsbA using two model organisms, P. aeruginosa and M. smegmatis. Our focus will be the study of substrates that are required for virulence and growth of these two microorganisms.
We seek to characterize the proteins involved in disulfide bond formation in Clostridium species and determine their role in toxin and virulence factor production. We will also investigate the impact of the bacterial and archaeal disulfide bond formation pathways in the gut microbiome by in situ identification of the proteins that are secreted and disulfide bonded.