Membrane protein efflux pump dynamics in biofilm formation and multidrug resistance

Membrane protein efflux pumps can expel a broad range of toxic substances out of bacteria which significantly contributes to multidrug resistance against functionally diverse antimicrobial drugs and biofilm formation. Biofilms typically pose a great issue for implants as they provide an ideal solid support to promote growth, thus treatment of such infections is extremely difficult, normally resulting in the removal of the implant.

Within this project we plan to expand our research in two ways: 1) the exploration of related efflux proteins and systems, away from the prototypical, to broaden our understanding of the fundamental role structural dynamics plays in multidrug resistance phenotypes, and 2) adapt our methods to interface with biofilm systems, to understand the role these efflux systems play in bacterial infection and resistance. To do this, microbiology, chemical biology, and advanced mass spectrometry strategies will be utilized and developed in unison to enable membrane protein dynamics to be deciphered within complex environments. Capturing these dynamics with Hydrogen/Deuterium eXchange Mass Spectrometry (HDX-MS), which measures the extent and rate of exchange of protein backbone amide hydrogens for deuterium. Combining these insights with biofilm systems and assays will provide unprecedented insight into the function of these systems, particularly on the impact of drug interactions and clinically relevant mutations.

We aim to uncover the structural dynamics of membrane proteins involved in antimicrobial resistance (AMR) of bacteria. AMR is recognised by the United Nations as a global health emergency. The projects’ focus is on multidrug efflux systems which play major roles in AMR, specifically the resistance-nodulation-division (RND) protein class.