BRET-based RAS biosensors that show a novel small molecule is an inhibitor of RAS-effector protein-protein interactions.
Nicolas BéryAbimael Cruz-MigoniCarole Jr BatailleCamilo E QuevedoHanna TulminAmi MillerAngela RussellSimon Ev PhillipsStephen B CarrTerence Howard RabbittsPublished in: eLife (2018)
The RAS family of proteins is amongst the most highly mutated in human cancers and has so far eluded drug therapy. Currently, much effort is being made to discover mutant RAS inhibitors and in vitro screening for RAS-binding drugs must be followed by cell-based assays. Here, we have developed a robust set of bioluminescence resonance energy transfer (BRET)-based RAS biosensors that enable monitoring of RAS-effector interaction inhibition in living cells. These include KRAS, HRAS and NRAS and a variety of different mutations that mirror those found in human cancers with the major RAS effectors such as CRAF, PI3K and RALGDS. We highlighted the utility of these RAS biosensors by showing a RAS-binding compound is a potent pan-RAS-effector interactions inhibitor in cells. The RAS biosensors represent a useful tool to investigate and characterize the potency of anti-RAS inhibitors in cells and more generally any RAS protein-protein interaction (PPI) in cells.
Keyphrases
- wild type
- small molecule
- energy transfer
- induced apoptosis
- protein protein
- emergency department
- endothelial cells
- mesenchymal stem cells
- stem cells
- dendritic cells
- immune response
- single cell
- cell cycle arrest
- high throughput
- oxidative stress
- cell therapy
- regulatory t cells
- fluorescent probe
- transcription factor
- induced pluripotent stem cells
- smoking cessation