NanoBRET TE Intracellular E3 Ligase Assay, VHL
Protein degradation via small molecules such as PROTACs or molecular glues is governed by multiple steps. The first step involves involves entry of the small molecule degrader compound into the cell, followed by binding to the E3 ubiquitin ligase and protein target. The NanoBRET TE Intracellular E3 Ligase Assays allow assessing both intracellular binding and availability. Specifically, the assays can be used to measure compound binding or affinity for an E3 ubiquitin ligase protein, Cereblon (CRBN) or von Hippel-Lindau disease tumor suppressor (VHL), in live cells.These assays are based on the NanoBRET technology, an energy transfer technique designed to measure molecular proximity in living cells. In addition, the assays can be used to quantify intracellular availability of the compound. As heterobifunctional degraders like PROTACs often have high molecular weight, they may suffer from poor cell permeability or intracellular availability. By performing the NanoBRET TE E3 Ligase Assay in two unique modes (live-cell and permeabilized-cell), the results can be used to calculate the intracellular availability of a degrader compound.