Table 2.
FRET-based diUb cleavage assay to measure deubiquitinating activity
| Enzyme | Activity with 1 μM inhibitor (% of uninhibited activity)a |
|---|---|
| STAMBP/STAM1+UbVSP.1 | 7.2 ± 0.6 |
| STAMBP/STAM1+UbVSP.3 | 13.4 ± 0.2 |
| STAMBPL1+UbVSP.1 | 44 ± 3 |
| STAMBPL1+UbVSP.3 | 118 ± 5 |
| STAMBPJAMM+UbVSP.1 | 0 |
| STAMBPJAMM+UbVSP.3 | 11 ± 2 |
| STAMBPJAMM+BC-1471 (CAS 896683-84-4) | 107 ± 2 |
| STAMBPJAMM+BC-1471 (CAS 896683-78-6) | 96 ± 3 |
a
Reactions were measured in triplicate at room temperature with 5 nM enzyme and 100 nM diUb K63 TAMRA substrate. Reported activity is normalized to identical reactions without added inhibitor.