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. Author manuscript; available in PMC: 2014 Jul 19.
Published in final edited form as: ACS Chem Biol. 2013 May 1;8(7):1435–1441. doi: 10.1021/cb400177q

Figure 1. Identification and characterization of a new family of SENP inhibitors.

Figure 1

(A) and (B) Representative Coomassie-stained gels showing cleavage of SUMO-1 and SUMO-2 by SENP1 and SENP2 in the presence of increasing concentrations of SPI-01 (A) and SPI-07 (B). YSE, fusion SUMO (S) precursors flanked by YFP (Y) and ECFP (E) at the N- and C-termini, respectively. (C) Effects of the panel of inhibitors (Table 1) at inhibiting SENP1, 2 and 7. In 96-well plates, SENPs (50–200 nM) were pre-treated with increasing concentrations of each compound, after which DUB-Glo (40 μM final concentration; Promega) was added as substrate. Experiments were performed in triplicate. The amount of cleaved product is proportional to the relative light unit (RLU), which is bioluminescence produced by luciferase catalyzed reaction of luciferin that was produced by SENP cleavage of DUB-Glo. (D) Accumulation of SUMO-2/3-modified proteins in HeLa cells upon treatment with increasing doses of SPI-01. (E) Retention of SUMOylated proteins during recovery of HeLa cells from heat shock in the presence of 60 μM SPI-01 and SPI-02.