One of the major challenges in phenotypic drug discovery programs is the need for target deconvolution and validation, identifying the specific proteins a drug interacts with. In collaboration with a client CETSA was used to address this issue through unbiased proteome-wide profiling in both intact cells and lysates.
The goal of this study was to identify the direct targets and elucidate the complete target profile and associated biology of a polyketide lactone. The objective was to identify its direct protein targets and map the full target profile and biological pathways affected—delivering deeper insights to progress the project.
The key questions for the study to answer was:
- Can we utilize CETSA in intact cells to determine the complete target profile and associated biology of the compound?
- Can we apply CETSA in lysate to differentiate between direct targets and downstream pathway events?
The collaboration successfully identified the direct targets, including GBF1, and provided valuable insights into its mechanism of action by comparing results from intact cells and lysates. The study revealed that it acts as a molecular glue, stabilizing the formation of an ARF1-GDP/GBF1 complex—a novel insight into its biological activity.
INSIGTHS: This case study highlights the power of CETSA in phenotypic drug discovery, offering a robust method for target deconvolution and validation without requiring modification of the drug or target. By staying true to the native biological context, CETSA enables deeper understanding and supports the continued advancement of drug discovery programs.
