New approaches to measuring single-molecule protein dynamics

Principal Advisor: Dr Sebastian Furness

Organisational unit: School of Biomedical Sciences

Research group: Furness Group - Laboratory of receptor transducer coupling

We recently discovered a new class of highly fluorescent zwitterionic fluorophores, derivatives of which display extreme environmentally sensitivity. These exciting molecules have physical properties that makes them interesting candidates for utility in a range of technologies. Of interest to us is the possibility that these molecules could be used to correlate conformational changes at G protein-coupled receptors (GPCR) which are an important class of drug targets. GPCRs are often activated by several different agonists, both endogenous and as medicines. The significance of this study is that differences in GPCR conformation are thought to be related to the ability of particular agonists to elicit cellular responses. This is because the cellular response is not simply a product of GPCR occupancy, as would be expected in an “on” – “off” switch or indeed to how tightly the agonist is bound. The project will involve the synthesis of novel fluorophores (Monash School of Chemistry), that will be subsequently conjugated with GPCRs or their signal transducers; G-Proteins. Interaction of these fluorescent, environmentally sensitive, GPCR or G-protein conjugates will then be investigated using single molecule dynamics with an emphasis on the utility of (Fluorescence Lifetime Imaging Microscopy – Fluorescence Resonance Energy Transfer) FLIM-FRET in addition to a suite of other techniques.