NanoBRET Assay Validates p53 Y220C Ligand Engagement in Living Cells
- Researchers built a live-cell NanoBRET assay to test whether reported p53 Y220C ligands truly engage the mutant protein inside intact cells.
- The BRET-based readout gives a cell-based engagement ranking for known Y220C stabilizer/reactivator compounds, complementing prior biochemical binding data.
- Such cellular validation checks permeability and intracellular target occupancy, a key hit-to-lead checkpoint for this hard-to-drug p53 cavity mutant.
💡 Points
Compounds that light up beautifully in DSF or ITC routinely stall out crossing a membrane into that buried, polar cavity — a cell-based BRET ranking is built to catch precisely that failure mode before a Y220C hit gets pushed into SAR. I'd still want it cross-referenced against a functional readout like p21 induction or apoptosis, and benchmarked directly against rezatapopt's (PC14586) cellular engagement window before trusting the ranking over biochemical Kd.
NanoBRET Target Engagement Enables Validation of p53 Y220C Ligands in a Cellular Context ↗