Molecular characterization of the interactions of the Resistance-Nodulation-Division Pump MexJK of Pseudomonas aeruginosa with outer membrane proteins OprM and OpmH.
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Pseudomonas aeruginosa is an important Gram-negative opportunistic pathogen, which has become a significant threat due to wide-spread antimicrobial resistance. Resistance Nodulation Division efflux pumps are important contributers to antimicrobial resistance in P. aeruginosa and other organisms. Resistance nodulation division efflux pumps are composed of three proteins; however, the mechanism by which the three proteins interact to form a functional complex remains, for the most part, unknown. The goal of this project is to further our understanding of how these proteins interact, to better understand how these efflux pumps operate in hopes of identifying novel therapeutic targets. We constructed a single copy expression system to study MexJK in the presence of either OprM or OpmH and confirmed these constructs using qRT, immunoblot and MIC. Chimeric OprM proteins were constructed by swapping α-helical domains from the OpmH protein. Using these chimeric proteins, we were able to identify regions within OprM/OpmH that may be responsible for substrate specificity by generating chimeric proteins. Taken together this data broadens our understanding of how this complex interacts.