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. 1997 Dec;8(12):2379–2390. doi: 10.1091/mbc.8.12.2379

Figure 5.

Figure 5

Characterization of the interactions of Ran and p97 with nucleoporins RanBP2 and p62. (A) Titration of Ran binding to RanBP2. Increasing concentrations of Ran loaded with either [32P]GTP or [32P]GDP were incubated with RanBP2 adsorbed to microtiter wells. After washing, the radioactivity was recovered from the wells with SDS and counted in a liquid scintillation counter, and the amount of Ran bound to RanBP2 was calculated from the [32P]GTP-specific activity. (B and C) Titration of p97 binding to RanBP2 and p62, respectively. Increasing concentration of His-p97 were incubated with RanBP2 (B) or p62 (C) and adsorbed to microtiter wells. The 6xHis-p97 that remained after washing was then cross-linked to the adsorbed protein and detected with antibodies (see MATERIALS AND METHODS). The Lineweaver-Burk plots used to determine the apparent binding constants (Kdapp.) are shown in the insets.