View All Posters

P2022
Detection of Ligand-Dependent Interaction of Nuclear Receptors with their Cofactor Using the Corning® Epic® System
Presenter Jack Fang, Corning Inc., USA
Additional Authors: Ingo Kober, L. Jack Fang, Johannes Gleitz, and Anthony G. Frutos
Cofactor recruitment is a crucial regulatory step for many nuclear receptors to regulate target gene expression. Ligand-binding of a nuclear receptor results in its conformational changes to interact and recruit cofactors that are critical for transcription activation. Small molecules that enhance or inhibit the interaction of a nuclear receptor with its cofactor will likely function as agonists and antagonists for the treatment of nuclear-receptor related diseases. Corning® Epic® System, a label-free, high-throughput detection technology, has been previously used for measuring direct binding of small molecule compounds to immobilized nuclear receptors. In this study, the Epic® System was used for detection and validation of small-molecule compound-dependent interactions between nuclear receptors and their common cofactor. The cofactor was directly immobilized via amine coupling in a 384-well Epic® plate; each nuclear receptor was premixed with a small molecule ligand and then added to the immobilized cofactor for direct binding detection. Both nuclear receptors were tested to determine the yes/no binding to the immobilized cofactor in the presence or absence of two Merck Serono ligands and the relative binding affinity (KD) of ligands to each receptor. The results presented here demonstrate that the Epic® System is able to not only detect the small molecule compound-dependent interaction between a nuclear receptor and its cofactor but also determine relative binding affinities in the presence of different small molecule compounds.