Figure 11.

Analysis of SUMO isopeptidase and pre-SUMO-processing activity of At5g60190. A, The SUMO isopeptidase model substrates AtSUMO1-ScPCNA, AtSUMO2-ScPCNA, and AtSUMO3-ScPCNA were analyzed before (−) and after incubation (+) with At5g60190. Proteins were separated on a 12% SDS-polyacrylamide gel, and SUMO-ScPCNA conjugates were detected by western-blot analysis using a Myc peptide-specific antiserum. B, The pre-SUMO-4CL reporter constructs were incubated with At5g60190 as depicted. As reference probes, AtSUMO1 (lane 1) and the untreated AtSUMO1-4CL protein (lane 2) were applied. Proteins were separated on a 15% SDS-polyacrylamide gel, and the pre-SUMO reporter constructs were detected by western-blot analysis using a HA peptide-specific antiserum.