Untagged RanY197A, GST-NESMVM (or mutants), GST-yRanBP1, and GST-yCRM1ΔH9 were expressed in E. coli separately and mixed, sonicated in lysis buffer (50 mM Tris pH 8.0, 200 mM NaCl,10% glycerol, 2 mM DTT, 5 mM MgCl2 and 1 mM PMSF). The complex was purified by GSH beads, cleaved off from beads by incubating the TEV enzyme overnight. The complex was further purified by gel filtration Superdex200 (GE Healthcare) column in buffer D (10 mM Tris 7.5, 100 mM NaCl, 5 mM Mg(OAc)2, 0.1 mM GTP, 2 mM BME). The complex was concentrated to 6 mg/mL using a Millipore spin concentrator (M.W. cutoff 10, 000). Crystals of different complexes were grown at room temperature by hanging drop vapor diffusion against 0.1 M Bis-Tris (pH 6.6), 0.2 M NH4NO3, and 18% PEG3350. Crystallization condition supplemented with glycerol (12% v/v) was used the as the cryoprotectant. X-ray diffraction data were collected at Shanghai Synchrotron Radiation Facility (SSRF) beamline BL17U1 and BL19U1.34 (link) Coordinates of yCRM1-hRan-yRanBP1 (PDB code: 4HAT) were used as the search model and refined with rigid body refinement briefly then restrained refinement using the program Refmac5.35 (link) Translation/Libration/Screw (TLS) refinement36 (link) was used in the refinement process. The data collection and refinement statistics are provided in Table S1.