Fig. 6

Fig. 6

Fig. 6. zUba7 prefers zISG15 and zUba1 prefers Ub. (A), Ub or zISG15 was titrated into reactions containing zUba1, ATP, and zISG15 or Ub fixed at 0.8 μM. (B), GST-Ub and GST-zISG15 titration. zUba7 was incubated with the indicated concentration of GST-UBL for 10 min at 30 °C. Samples were immunoblotted with the anti-Flag. (C), Time course for GST-Ub and GST-zISG15 activation. Reactions containing 1.2 μm GST-UBL and zUba7 were incubated for the indicated time at 30 °C. Samples were immunoblotted with the anti-Flag. (D) Ub or zISG15 was titrated into reactions containing zUba7, ATP, and zISG15 or Ub fixed at 0.8 μM. (E) Kinetics of Ub activation by zUba1 and zUba7. (F) zUba1 or zUba7 was titrated into reactions containing Ub, ATP, and zUba7 or zUba1 fixed at 2 μM. (G) Myc-Ub titration: 2 μM zUba1 or zUba7 was incubated with the indicated concentration of Myc-Ub for 10 min at 30 °C. The samples were immunoblotted with anti-Myc.