Figure 7.

The ability of RanGTP to modulate the binding of p97 to RanBP2 and p62 requires the RanGTP/p97 interaction. (A) 3 nM or 24 nM [³²P]GTP-Ran was incubated with wt p97 or ΔN p97 bound to microtiter wells. After binding and washing, the radioactivity was recovered from the plate by elution with SDS and was counted in a liquid scintillation counter. The amount of bound RanGTP was calculated from the [³²P]RanGTP-specific activity. (B) The binding of wt p97 and ΔN p97 to RanBP2 adsorbed to microtiter wells was analyzed in the absence of Ran, or in the presence of 3 nM or 24 nM RanGTP, and the bound p97 was quantified as in Figure 4. (C) The binding of wt p97 and ΔN p97 to p62 adsorbed to microtiter wells was analyzed as in panel B.