Figure 6

In vitro and in vivo cleavage of His6SUMO-eDAL by the Ulp1 constructs. A SDS-PAGE analysis of the purified fusion protein and two Ulp1 constructs (His6MBP-Ulp1 and His6-Ulp1). The corresponding proteins are indicated by the arrows. B: in vitro cleavage of SUMO-eDAL by two Ulp1 constructs at the different temperatures by coupled assay. C-E: in vivo cleavage of SUMO-eDAL by Ulp1 constructs detected by SDS-PAGE and Western blot analysis, and activity measurement of the released DAL. Lane1: cells carrying the plasmids pSUMO-eDAL and pMAL-c2x. Lane 2: cells containing pSUMO-eDAL and pMAL-c2x-Ulp1. Lane 3: cells harboring pSUMO-eDAL and pCDF-Duet1. Lane 4: cells carrying pSUMO-eDAL and pCDF-Ulp1. The expressed His6SUMO-eDAL and cleaved His6SUMO were indicated by the arrows. The colume in Figure 6E represented the corresponding DAL activities. The in vivo and in vitro activity of DAL as the fusion partner was applied as the control and subtracted.