FIG. 5.
UCH-L3 labeling by 125I-Ub-VS and 125I-Nedd8-VS. 125I-Ub-VS (2.5 × 105 cpm) (A) or 125I-Nedd8-VS (2.5 × 105 cpm) (B) was incubated for 1 h at 37°C with lysate buffer (first lane), 20 μg of EL-4 lysate, or 150 ng of recombinant, purified UCH-L3. The reactions were terminated as described for Fig. 2. Polypeptides were resolved by SDS-11% PAGE and visualized by autoradiography. The triangles mark the UCH-L3-Ub-VS (left) and UCH-L3-Nedd8-VS (right) adducts. The positions of molecular mass markers (in kilodaltons) are indicated on the left.