Identification of active deubiquitinases in dendritic cells under unstimulated and activated conditions

Abstract

Ubiquitination is a post-translational modification with well described roles in the regulation of immune cell function, such as dendritic cells (DCs). DCs are crucial initiators and regulators of T cell-mediated immune responses. In this study, we use activity-based probes and proteomics to systematically profile DUBs in a DC line. Using the lysate-based biotin(bio)-Ubiquitin(Ub)-propargylamide(Prg) probe and proteomics, the activity and abundance of deubiquitinases was measured in unstimulated and CpG-stimulated MutuDCs to simulate responses to bacteria. This analysis was complemented with profiling using the cell-permeable DUB activity-based probe IMP-2373. The combined analysis with bio-Ub-Prg and IMP-2373 provided insights into the dynamics of DUB activity and abundance in DCs following stimulation and identified several DUBs for further study.