Research

Stochastic detection of protein kinases

I have developed a label-free, single-molecule method for the detection of protein kinases. Using a novel protein engineering strategy, I produced alpha-hemolysin nanopores bearing kinase substrate peptides attached by a single terminus to the pore mouth.

When a single pore is inserted into an artificial lipid membranes, the peptide substrate protrudes into solution, and the binding and dissociation of kinase molecules can be observed in real time through the modulation of ionic current through the pore under an applied potential.

I have applied this method to understand the mechanism of substrate binding for the Pim family of kinases, revealing electrostatically enhanced association. Further, I have shown that these engineered nanopore sensors can be used for label-free inhibitor screening, with the potential for high-throughput scaling.

Engineering biological pattern formation

Biological self-organisation through spontaneous symmetry breaking is essential for the emergence and maintenance of life. Important examples include cellular polarisation, and the waves of E. coli Min proteins that identify the mid-cell for division.

I aim to design and engineer artificial systems mimicking biological pattern formation to test proposed mechanisms, and for use as building-blocks in higher-level mimics of life such as protocells.