FIGURE 2.

Involvement of the switch II region of Rab35 in the binding of Rab35BPs. A, sequence alignment of the switch II regions of mouse Rab5A and Rab35. Amino acid residues that are conserved in the two sequences are shown against a black background. The central parts of their switch II region (dashed line) were swapped, and the Rab35(S5A) mutant contains part of the switch II region of Rab5A. B, the switch II region of Rab35 is required for binding all Rab35BPs except RUSC2. Yeast two-hybrid assays were performed as described in the legend for Fig. 1C. C, the Rab35(S5A) mutant hardly interacted with centaurin-β2 at all in contrast to the weak centaurin-β2 binding activity of the Rab35(T76S/T81A) mutant. Co-immunoprecipitation assays were performed as described in the legend for Fig. 1E. The positions of the molecular mass markers (in kilodaltons) are shown on the left. IP, immunoprecipitation.