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The C-terminal SIM of Ulp2 facilitates its cleavage of in vitro synthesized poly-SUMO chains as well as native sumoylated proteins. A, in vitro sumoylation to generate poly-SUMO chains. B, cleavage kinetics of in vitro synthesized poly-SUMO chains by Ulp2400–767 enzyme (WT and SIM3A). C, purification of sumoylated proteins from the ulp2Δ mutant using anti-FLAG affinity resins. D, cleavage kinetics of native sumoylated proteins by Ulp2400–767 enzyme (WT and SIM3A).
