Networks of protein-protein interactions are central to cell and developmental biology and of great importance in molecular medicine. Given the potential importance of protein-protein interactions to healthcare, wealth creation and the biotech industry the re is an urgent need to train young researchers in the latest technologies aimed at identifying and dissecting such interactions by the very latest in high-throughput approaches, bio-informatics and rigorous biophysical analysis. We will train early-stage and experienced researchers to use new bio-informatics tools to direct high-throughput screening (HTS) for new protein-protein interactions. Network "hubs" or "nodes" (i.e. proteins/protein families that interact crucially with one or many other partners in the network) will be identified and used as baits in HTS.
Based on previous collaborative, published work, we will develop an in vitro selection system for protein-protein interactions by in vitro compartmentalisation (IVC). IVC relies on dispersing aqueous reactions into microscopic water-in-oil emulsion droplets ( ~5 fl) enabling selection of 10e10 pairs of interacting proteins. Hence, all ~10e9 possible pair wise binding interaction of proteins encoded by the human genome are possible in a more economical format than existing HTS techniques.
A novel system for covalently labelling proteins will be used to label and cross-link proteins in vivo and to create protein micro-arrays. Kinetic and thermodynamic characterisation of identified protein-protein interact ions will be performed using a variety of techniques including a novel acoustic biosensor. Most importantly, we aim at an integrated approach where the bio-informatics tools we develop will be used to direct the HTS efforts. In turn, results obtained from t he screening will help validate and tune our newly-developed computational tools, while detailed mechanistic studies will provide insight into the physiological role of newly-identified interacting pairs.
Call for proposal
See other projects for this call