About the Project
The interface between the actin cytoskeleton and the plasma membrane is a master regulator of cell shape, rigidity, and motility. These critical structures in the cell are linked together by proteins in the ERM protein family, which connect lipids in the plasma membrane to underlying actin filaments. Regulation of cytoskeleton-membrane interactions is important for proper cell function, helping to tell the cell when and where to move. Dysregulation of this system can cause immune dysfunction, loss of tissue structure, and spread of diseased cells. For example, dysregulation of the ERM protein ezrin drives tumor progression and metastatic spread in cancer. Given its importance, selectively manipulating the actin-plasma membrane interaction is of great interest to control the fate of cells in health and disease. While attachment of ERM proteins to the plasma membrane is well-characterised, the details of how they bind actin is unknown. Recent studies point to two amino acids in the C-terminus, but high-resolution structural data is missing for this region leaving a gap in our understanding of this system and hampering our ability to exploit it in the development of therapeutics. This is illustrated by the wholescale failure of small-molecule inhibitors targeting the C-terminal region to make it to clinic for treatment of cancer. A structural understanding would not only complete our picture of this critical system but would open clinical opportunities that do not currently exist. We will characterise the structure of the C-terminal domain of an ERM protein, its binding to actin filaments and the effect of novel, structure-led actin binding inhibitors developed during the project. You will be using a combination of techniques in molecular biology (protein expression and purification), biochemistry (electrophoresis, PCR) and biophysics (NMR, advanced fluorescence microscopy) and perform experiments on live cells as well as minimal, reconstituted actin/membrane systems.
