Figure 3

Release of the (A) Strep and (B) FLS peptides from Ecotin-Ubiquitin fusion proteins by UCH-L3. Expression and cleavage of the fusion proteins in the periplasmic extract was analyzed by SDS-PAGE and Strep-Tactin^® HRP immunoblots. M: protein marker, -I and +I: before and after induction of expression, IB: inclusion bodies, Cyt: cytoplasm, Per: 1 × and 5×, single and fivefold amount of periplasmic protein, c1: periplasmic extract untreated, c2: periplamic extract incubated in UCH buffer, *: UCH-L3 digested periplasm, CS: culture supernatant. The upper arrow indicates intact fusion protein, the lower one with the asterisk the major cleavage moiety Ecotin-Ubiquitin (24.51 kDa).