Figure 2. mRanBP1’s RBD dissociates CRM1 cargo through sequestering RanGTP.

(A–D) GST-NES^PKI pull down of CRM1 and RanGTP in the presence of RanBP1 or its mutants. (A) NESmut or ΔC dissociates cargo as WT RanBP1. (B) High concentration of RanGTP inhibits cargo dissociation in animals but not yeast. (C) RanGTP-bound ΔC loses its ability in cargo dissociation while yeast RanGTP-RanBP1 remains competent. (D) ΔC dissociates cargo when its concentration is higher than that of RanGTP, and yRanBP1 dissociates cargo when its concentration is higher than that of CRM1.

Figure 2—figure supplement 1. GST-NES pull down shows that NESmut or ΔC is as potent as WT RanBP1 in dissociating CRM1 complex.

There is no obvious cargo dissociation difference among RanBP1 constructs for each concentration tested.