FIGURE 3. Structural analysis of the complete cargo-recognition complex.

a, Alignment of human VPS35 repeats. Columns of residues that are likely to form the hydrophobic core of the structure have a yellow background. Apolar residues are in red. Cylinders show the -helical regions. Magenta boxes within the -helices indicate the positions of apolar residues in the consensus sequences. The C-terminal region of the crystal structure is in bold. Loop regions predicted on the basis of the multiple sequence alignment of VPS35 proteins are shown by lower-case letters. b, Averaged images of the cargo-recognition complex from negative stain electron microscopy (Supplementary Fig. 6). The images were obtained by multivariate statistical analysis with reference-free alignment. The number of images per class 1 to 5 is respectively 288, 362, 367, 206 and 327. c, Corresponding projections, limited to 25 Å resolution, of the cargo-recognition complex model shown in d (C-terminal crystallized region of VPS35, red; VPS29, green; VPS26, cyan; and N-terminal modelled region of VPS35, orange). Each image in c is oriented such that VPS35–VPS29 corresponds always to the top part of the image and VPS26 to the bottom part, as in d. The correlation coefficient between the electron microscopy class average and the corresponding model projection for class 1 to 5 is respectively 0.79, 0.70, 0.86, 0.75 and 0.73. Scale bar in c, 100 Å.