Measurement of PROTAC ternary complex formation using the switchSENSE Y-structure and FRET signals

Abstract

Proteolysis targeting chimeras (PROTACs) are essential bifunctional small molecules that engage the formation of a ternary complex consisting of an E3 ubiquitin ligase, a target protein of interest and the PROTAC itself. Using switchSENSE® technology and the novel DNA Y-structure, an E3 ligase as well as a target protein can be functionalized on each separate end of two FRET pair color-coded Y-arms. The Y-structure closes upon PROTAC binding and the subsequent ternary complex formation bringing together the green donor and the red acceptor dye into a closer, FRET sensitive, distance.