The control of numerous bacterial diseases and infections through the use of antibiotics has arguably been one of the greatest achievements of modern medical science. However, the inexorable rise in antibiotic resistance among pathogens poses an increasing threat, with the very real possibility that previously easily-controlled infections might again become untreatable diseases.
One key mechanism through which bacteria may become resistant to multiple antibiotics involves the up-regulation of multidrug (MDR) efflux pumps in the bacterial cell envelope which eject drugs and other noxious molecules, including antibiotics, out of bacterial cells. Our laboratory studies the structure and operation of these important biological machines at the molecular level, opening the possibility that we might eventually develop countermeasures to antimicrobial drug resistance.
This project is of a molecular/biochemical nature. You will seek to purify and structurally characterise a novel family of efflux pumps identified in Clostridium difficile, the cause of pseudomembranous colitis and a prominent hospital acquired infection. The ultimate aim is to determine the atomic structure of MDR pumps by X-ray crystallography.
Working on this project, you will gain laboratory experience in cloning, protein expression and purification, ITC (isothermal titration calorimetry) and X-ray crystallography - all established techniques within our lab.
- Koronakis V, Sharff A, Koronakis E, Luisi B, Hughes C (2000) Crystal structure of the bacterial membrane protein TolC central to multidrug efflux and protein export. Nature 405: 914 - 919
- Andersen C, Koronakis E, Bokma E, Eswaran J, Humphreys D, Hughes C, Koronakis V (2002). Transition to the open state of the TolC periplasmic tunnel entrance. Proceedings of the National Academy of Sciences (USA) 99: 11103 - 11108
- Higgins M, Boekma E, Koronakis E, Hughes C, Koronakis V (2004) Structure of the periplasmic component of a bacterial antibiotic efflux pump. Proceedings of the National Academy of Sciences (USA) 101: 9994-99