Figure 4.

Cleavage of H862Q-containing mutants. Purified Nup98(H862Q) and Nup98(H862Q/S864C) are in lanes 1 and 2. Nup98(H862Q/S864C) was incubated at ambient temperature for 20 hr with 25, 50, and 200 mM DTT (lanes 3, 4, and 5, respectively). SDS-denatured and native Nup98(H862Q/S864C) were incubated with 200 mM NH₂OH at room temperature for 15 min (lanes 6 and 7) and 15 hr (lanes 8 and 9). Additionally, the contrast of the lower region of lanes 3–9 was enhanced (Lower) to facilitate detection of Nup98-C. Proteins were separated by SDS/4–12% NuPAGE (NOVEX, San Diego) and visualized with Coomassie blue R-250.