Crystallization of the Catalytic Domain of Bacterial PhoQ for Rational Design of Inhibitors
Worldwide, about 4.95 million deaths are attributed to antimicrobial resistant infections, resulting from antibiotic overuse and the lack of novel antibiotic development. One alternative treatment targets inhibiting bacterial virulence mechanisms, which are what cause harm in the infected host. Two-component systems (TCS) are bacterial signaling pathways that are often involved in controlling virulence. Previous work used small molecule libraries to identify potential scaffolds for inhibition of histidine kinases within the TCS. Using structural knowledge of the catalytic (CA) and ATP binding domain of the histidine kinase PhoQ from Salmonella enterica, we plan to rationally design novel small molecules inhibitors to hinder bacterial virulence. This will be accomplished through cloning the DNA sequence of the PhoQ CA domain from S. enterica and inserting it into a recombinant overexpression plasmid. By inserting this plasmid into E. coli, a protein containing the CA domain of PhoQ will be produced and we can then purify and crystallize this biomolecule. Finally, through use of protein crystallography with inhibitor scaffolds and structural binding knowledge, novel small molecule inhibitors will be developed.