Matt Robers1, Mei Cong1, Pete Stecha1, Brock Binkowski1, Christopher Eggers1, Danette Daniels1, Marie Schwinn1, Braeden Butler1, Thomas Machleidt1, Mary Hall1, James Unch1, Kris Zimmerman1, Paul Otto1, Gedi Vidugiris1, Monika Wood1, Mike Slater1, Suzanne Grooby2, Holly Astley2, Chris Lowe2, Lance Encell1, Frank Fan1 and Keith Wood1
1Promega Corporation, 2800 Woods Hollow Road, Madison, WI 53711 USA; 2Horizon Discovery Ltd, Building 7100, Cambridge Research Park, Cambridge, CB25 9TL, United Kingdom
The newly developed NanoLuc® (Nluc) luciferase is a small (19.1kDa) engineered luminescent reporter. The enzyme is about 150-fold brighter than either firefly (Photinus pyralis) or Renilla reniformis luciferase, using a novel coelenterazine analog (furimazine) to produce high intensity, glow-type luminescence. The small and bright features of Nluc enable the development of homogenous assays reporting on pathway analysis via endogenous promoter or protein, G protein coupled receptor (GPCR) and Receptor Tyrosine kinase (RTK) endocytosis, small molecule-protein interactions using Bioluminescence Resonance Energy Transfer (BRET). Furthermore, Protein–protein interactions (PPI) have traditionally been targeted by many drug developers due to their therapeutic relevance. Using Nluc as donor and HaloTag® as acceptor, we were able to monitor protein-protein interactions to monitor chromatin interactions.