This research project with Dr. Adam Hoppe at South Dakota State University (SDSU) focused on improving Fluorescent Resonance Energy Transfer (FRET) efficiency.  FRET is a tool used in biology to measure the interaction of proteins by attaching donor and acceptor fluorescent proteins (FP) to two potentially interacting proteins.  When a specific wavelength of light is shined on the donor FP it gives off light in the form of fluorescence.  If however, the two tagged proteins interact bringing the donor and acceptor

FPs to within about 5 nm of each other, the donor can transfer its energy to the acceptor FP which in turn fluoresces at a different color than the donor.  By measuring the appearance of this FRET signal, the interaction of the two proteins can be measured.  However, FRET is limited in its ability to detect interactions between large (>25 kDa) proteins since the donor and acceptor FPs often do not reach the required 5 nm separation distance.  This research project aimed to overcome this limitation by attaching low affinity peptide motifs to the donor and acceptor FPs. Interactions between these motifs are expected to bring the FPs to within the 5nm distance needed for efficient FRET. If successful, the outcome of this research will be new tools that enable detection of protein interactions by FRET over a much larger range of protein sizes than previously possible.

 

Keary Johnson loading a gel

 

 

 

 

Students examining fluorescent proteins