FIGURE 3.

Ulp2 does not discriminate between full-length Smt3 and N-terminally truncated (ΔN17) Smt3. Comparison of band patterns derived from Ulp2-mediated processing of substrates harboring Smt3 chains led either by full-length (FL) Smt3 or by N-terminally truncated (ΔN17) Smt3. These 5xSmt3-GFP substrates were incubated with E. coli lysates containing Ulp1 or Ulp2 diluted in activity test buffer (1:1, undiluted lysate). As a control, a lysate was used that contained the inactive C624A variant of Ulp2. Substrates were incubated with different dilutions of Ulp2-containing lysates for 2 h at 30 °C. Reaction products were then analyzed by SDS-PAGE and anti-HA Western blotting. inact., inactive.