Studying OTUD6B-OTUB1 Protein–Protein Interaction by Low-Throughput GFP-Trap Assays and High-Throughput AlphaScreen Assays

Protein-protein interactions (PPI) are involved in a myriad of cellular processes, and their deregulation can lead to many diseases. One such process is protein ubiquitination that requires an orchestrated action three key enzymes add ubiquitin moieties substrate proteins. Importantly, this reversible through deubiquitinating enzymes. Both deubiquitination require PPIs once classified be utilized identify small molecule inhibitors counteracting these reactions. Here, we study the protein-protein interaction between two OTUB1 OTUD6B report for first time both proteins directly interact with each other. We describe GFP-Trap immunoprecipitation as cell-based method analyze OTUD6B-OTUB1 context AlphaScreen (amplified luminescent proximity homogeneous assay) assay tool detect direct search PPI inhibitors.