DUB activity-based probes

DUB activity-based probes (also known as ubiquitin activity-based probes or ubiquitin probes) are specialized tools designed to investigate the activity of deubiquitinating enzymes (DUBs) with high specificity and sensitivity. They are based on a ubiquitin or ubiquitin-like protein modified with a C-terminal reactive group (or warhead) that covalently modifies the active site of a target enzyme in a highly selective manner. Activity-based probes have proven to be critical research tools for advancing our understanding of ubiquitin biology by enabling researchers not only to track and analyze the activity of DUBs but also to study their structural details. Thus, by integrating these probes into their studies, scientists can uncover how DUBs malfunction and potentially link this to various diseases, providing a foundation for targeted therapeutic interventions. The addition of fluorescent dyes and/or affinity tags (such as biotin) allows labeled proteins to be visualized and enriched for subsequent studies.