Deubiquitinating enzymes (DUBs) are proteases that reverse ubiquitin modifications and can rescue a protein from proteasomal degradation. They specifically cleave ubiquitin-linked molecules downstream of the terminal carbonyl of the last residue of ubiquitin (Gly76). DUBs are classified into five families. Well-defined families include ubiquitin carboxyl-terminal hydrolases (UCH) and ubiquitin-processing proteases (UBP). They act as regulators of diverse cellular processes, such as development, growth and DNA repair. DUBs have become actively studied molecular targets for drug discovery since they are implicated in the pathogenesis of many human diseases.
The SensoLyte® 440 Deubiquitination Assay Kit provides a convenient assay for the screening of enzyme inhibitors and activators or for continuous assay of deubiquitination activity. A natural substrate, ubiquitin, coupled with the AMC (7-amino-4-methylcoumarin) fluorophore is used in the assay. Upon the cleavage by a deubiquitin protease, this substrate generates the blue fluorophore that can be detected at excitation/emission=354/442 nm.