Fig. 2: ScaC binds the C-terminus of BICD2.

A Chromatogram for the BICD2-ScaC complex (black). Purified BICD2 and ScaC were mixed in 1:2 molar ratio. Individual traces for BICD2 (orange) and ScaC (green) are shown. SDS-PAGE gels show fractions underlying the relevant protein peaks. This SEC experiment was repeated three times (B) SEC-MALS for the BICD2-ScaC complex. The molar mass of the complex is indicated. C Schematic diagram showing mapping of the ScaC-binding domain (highlighted in grey) using BICD2 truncations. D Crosslinking/MS of purified BICD2-ScaC complexes using ECD. Heteromeric crosslinks are shown. E Chromatogram of the BICD2-ScaC^59-159 complex. The SDS-PAGE gel shows fractions underlying the complex peak. This SEC experiment was repeated three times (F) AlphaFold prediction of BICD2^711-800. The residues involved in binding to RANBP2 and RAB6³⁵ are highlighted in pink and green respectively. G SDS-PAGE gels showing in vitro pull-downs of different Strep-BICD2^711-800 mutants in the presence of 3.5-fold excess GCN4-RANBP2^min, RAB6^Q72L or ScaC. This pulldown experiment was repeated three times (H) Competition pull-down assay using Strep-BICD2^711-800 in the presence of different combinations of GCN4-RANBP2^min, RAB6^Q72L and ScaC-SNAP. I Quantification of GCN4-RANBP2^min (top) and RAB6^Q72L (bottom) binding to Strep-BICD2^711-800 in the absence or presence of ScaC-SNAP. Error bars indicate mean ± standard deviation. P-values were calculated from n = 3 independent replicates using a two-tailed T-test. **p < 0.01. RANBP2 p = 8.8 × 10^-5. RAB6 p = 0.0025. Source data are provided as a Source Data file.