Quantifying CDK inhibitor selectivity in live cells.
Carrow I WellsJames D VastaCesear R CoronaJennifer WilkinsonChad A ZimprichMorgan R IngoldJulie E PickettDavid Harold DrewryKathryn M PughMarie K SchwinnByounghoon Brian HwangHicham ZegzoutiKilian V M HuberMei CongPoncho L MeisenheimerTimothy M WillsonMatthew B RobersPublished in: Nature communications (2020)
Concerted multidisciplinary efforts have led to the development of Cyclin-Dependent Kinase inhibitors (CDKi's) as small molecule drugs and chemical probes of intracellular CDK function. However, conflicting data has been reported on the inhibitory potency of CDKi's and a systematic characterization of affinity and selectivity against intracellular CDKs is lacking. We have developed a panel of cell-permeable energy transfer probes to quantify target occupancy for all 21 human CDKs in live cells, and present a comprehensive evaluation of intracellular isozyme potency and selectivity for a collection of 46 clinically-advanced CDKi's and tool molecules. We observed unexpected intracellular activity profiles for a number of CDKi's, offering avenues for repurposing of highly potent molecules as probes for previously unreported targets. Overall, we provide a broadly applicable method for evaluating the selectivity of CDK inhibitors in living cells, and present a refined set of tool molecules to study CDK function.
Keyphrases
- living cells
- small molecule
- cell cycle
- fluorescent probe
- induced apoptosis
- cell cycle arrest
- single molecule
- energy transfer
- reactive oxygen species
- endothelial cells
- protein protein
- cell proliferation
- oxidative stress
- single cell
- structural basis
- signaling pathway
- pi k akt
- quantum dots
- mesenchymal stem cells
- induced pluripotent stem cells
- stem cells
- data analysis