(
A) Coomassie staining of recombinant hinge-LBD mSF-1 protein (aa178-462) with DMSO (0) or with 10 µM TA. This protein fragment of mSF-1 contains only one of the two conserved sumoylation consensus sites at K194. Migration of SUMO1-SF-1 and unmodified SF-1 hinge-LBD (SF-1) are indicated by arrows. (
B) IVS of full length IκBα without (0) or with increasing concentrations of TA, as indicated, with SUMO-IκBα and unmodified IκBα indicated by arrows. (
C) Dose-dependent inhibition of IVS of fluorescently labeled AR peptide by TA with the IC
50 provided. Data are plotted as percent conversion versus TA concentration (log
10 [µM], left panel). Real-time sumoylation of AR peptide (% Conversion) are plotted at different concentrations of TA (right panel). IVS conditions and detection of AR peptide sumoylation by electrophoretic mobility shift assay are previously described in
Kim et al., (2013).
IVS: In vitro sumoylation; TA: Tannic acid.