FRET@ the short end: the development and application of single-molecule fluorescence techniques

Single-molecule Biophysics Lab – University of Sheffield

An exciting opportunity has arisen to join the Single-molecule Biophysics Lab at the University of Sheffield. Research in the lab involves the development and application of single-molecule fluorescence techniques to addressing crucial questions across physics, chemistry and the life sciences. You will join a multidisciplinary environment, which benefits from many national and international collaborations (Florida, Madrid, Bristol, Leeds). One project is detailed below but other projects are also available.

Project title: FRET@ the short end

Single-molecule Förster Resonance Energy Transfer (smFRET) is a powerful tool for monitoring distances between 40-100 Å. However, there is a need to extend the sensitivity to shorter distances which would enable the study of important conformational changes in smaller proteins and de novo protein structure determination by FRET. The dynamic range of a dye pair is characterized by the Förster radius (R0), defined as the inter-dye distance at which the FRET efficiency is 50%. Popular single-molecule dye pairs have Förster radii in the 50–65 Å range, ruling them out for detecting shorter distances (20-40 Å). This project will involve characterising a range of FRET pairs and linker chemistries to develop both ensemble and single-molecule assays for measuring these shorter distances.

Once established, these novel methods will be applied to a range of different systems, including protein-protein interactions in synthetic biology, assay development for allosteric drug discovery, and de novo protein structure determination.

Skills: The successful candidate will receive full training in a wide range of biochemical and biophysical techniques, chiefly single-molecule FRET spectroscopy and biomolecular labelling. Skills may also be gained by working closely with our collaborators, with significant scope to develop the project according to individual interests.