Investigating bacterial transporters that convey antibiotic resistance (360G-Wellcome-207152_Z_17_Z)

The rise of antimicrobial resistance (AMR) threatens many of the major advances in modern healthcare as the treatments rely on the implicit ability of the antibiotic to allow the body to heal from a serious infection. Antibiotic efflux transport is central to the development of AMR. At the single cell level, it is becoming apparent that the use of efflux pumps is the first line of defence against an antibiotic. These pumps decrease the intracellular level of antibiotic while the cell activates the various other levels of protection. This frontline of defence involves a coordinated network of efflux transporters. In the future, inhibition of this efflux transporter network, as a target for novel antibiotic therapy, will require the isolation and then biochemical/biophysical characterisation of each pump against all known and new antibiotics. This depth of knowledge is required so that we can fully understand and tackle the mechanisms of developing antimicrobial resistance. The key goals of this project are to over-express, IMAC purify a hexahistidine-tagged versions of a number of the efflux transporter and then characterise ligand binding against a set of clinically relevant antibiotics. The method used to measure ligand binding is thermophoresis (Nonotemper, Monolith).