Figure 1.

AO7 is involved in E2-dependent ubiquitination in vitro. (A) Equimolar amounts of the indicated GS-bound fusion proteins or GST alone were incubated in the presence of E1, E2 (UbcH5B), proteins from bacterial lysates, and ³²P-labeled Ub for 1.5 h at 30°C. The entire reaction was resolved under reducing conditions by SDS/PAGE. (B) GST fusion proteins were incubated as described for A in the absence or presence of E2 (UbcH5B). GST fusions of AO7T, Nedd4, and E6-AP migrate at approximately 69, 120, and 115 kDa, respectively.