Biophotonics

While FRET is often used visually in vivo, it may also be used as a quantitative screening assay for protein interaction. In this work, we study the small ubiquitin-like modifier SUMO1 and its interactions with further proteins of the SUMO cycle, including enzymes (Ubc9), substrate (RanGAP1) and proteases (SENP1), which are important in many biological processes. In this example, complexes of CFP-SUMO and YFP-Ubc9 were expressed and purified, and steady state FRET measurements of the binding of Ubc9 to SUMO1 were performed. This provides a real-time conjugation and protease cleaving assay, with a sensitivity and simplicity novel to biomolecular science. 

Key References

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