LiP-MS on HEK293 lysate treated with Probe Candidate TR05481366 and Negative Control TR05302201

Limited proteolysis coupled to mass spectrometry (LiP-MS) is a structural proteomics technique that can be used to identify the targets of small molecules. This is achieved by incubating lysate with the drug of interest, which induces structural changes such as occupation of the binding site or changes and protein-protein interactions. This is followed by a limited proteolysis step in which the unspecific protease proteinase K is added to the treated lysate. The protease preferentially cleaves accessible and flexible regions of the proteins, thus generating peptides that reflect the changes in surface accessibility caused by the treatment with the small molecule. Here we treated HEK293 cell lysate with the candidate probe TR05481366 and the corresponding negative control TR05302201 to profile target engagement and other structural effects. This project is part of the IMI EUbOPEN project and was performed in collaboration with Kilian Huber (University of Oxford).