Identification of deubiquitinases controlled by extracellular matrix stiffness in cancer cells

The project was aimed at identifying deubiquitinases (DUBs) involved in cancer cell mechanosignaling. Lysates from cancer cells grown on collagen matrices with varying stiffnesses were incubated with an activity-based HA-tagged ubiquitin probe that covalently binds DUB active site. To identify DUBs differentially activated on soft and stiff collagen hydrogels, lysates were immunoprecipitated with anti-HA agarose beads. Immunoprecipitates were further identified by LC-MS/MS analysis.