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. Author manuscript; available in PMC: 2019 Nov 20.
Published in final edited form as: Biochemistry. 2018 Nov 5;57(46):6538–6550. doi: 10.1021/acs.biochem.8b00987

Figure 2.

Figure 2.

BicD2/cargo complexes predominantly form 2:2 oligomers.(A) The purified BicD2-CTD was analyzed by SEC-MALS (at 2 mg/mL). Rayleigh ratio (blue) and molar mass MW (red) versus the elution volume are shown. The determined molar mass is MW = 23.3 ± 1.3 kDa, which closely matches the mass of a BicD2 dimer (20.8 kDa). (B) Purified Nup358-RBD2/BicD2-CTD complex was analyzed by SEC-MALS (at 14 mg/mL). Refractive index (blue) and molar mass MW (red) versus the elution volume are shown. The determined molar mass is MW = 155.4 ± 7.8 kDa, which is slightly below the molar mass of a 2:2 complex of Nup358-RBD2/BicD2-CTD (174.6 kDa). (C) The purified Rab6GTPmin was analyzed by SEC-MALS (at a concentration of 6 mg/mL). Rayleigh ratio (blue) and molar mass MW (red) versus the elution volume are shown. The determined molar mass is MW = 21.5 ±1.0 kDa, closely matching the molar mass of a Rab6GTPmin monomer (19.0 kDa). (D) The Rab6GTP/BicD2-CTD complex was analyzed at a protein concentration of 7 mg/mL by SEC-MALS. The molar mass of the major peak is MW= 61.0 ± 3 kDa, which is slightly less than the molar mass of a 2:2 complex (69.2 kDa).