Figure 1.
Schematic overview of full-length p115.
The construct comprising p115GHR used for crystallization is shown in gray.
Figure 2.
(A) Stereo view of the overall structure. The protein is composed of 11 armadillo repeats. (B) Structure-based sequence alignment of the p115 armadillo repeats and the loop regions. Consensus residues that define the conserved hydrophobic residues of each armadillo repeat are highlighted in blue; amino acids that define conserved polar, neutral residues are highlighted in green. Glycine and proline residues are highlighted in brown and olive, respectively. The repeat numbers are shown on the left. The sequences that form helices H1, H2, and H3 are indicated as green, blue and yellow cylinders. The corresponding ARM loops are marked on top, the USO helix is indicated as red cylinder. (C) N-terminal armadillo-like helical domain (left) and C-terminal Uso1 head domain (right) of p115GHR, elongated loops with at least 5 residues are colored in red (H1 green, H2 blue, H3 yellow).
Figure 3.
Interaction of p115GHR and the COG complex subunit COG2.
(A) Electrostatic surfaces of p115GHR. The amino terminus of the molecule is at the top of the figure. Blue indicates positive charge and red negative charge at the level of 10 kT/e. (B) Conserved residues of p115 and the yeast homolog Uso1p form a highly charged surface of exposed helices which define the COG2 binding site.
Figure 4.
Dimeric arrangement of the p115 globular head region.
(A) B-factor representation (“S-“ and “W-view”) of p115GHR molecules aligned by crystal symmetry. The intermolecular contact area (blue) is among the most rigid parts of the structure. (B) Depending on the orientation, the crystallographic dimer of p115GHR has a single-head (“O-view”) or double-lobed globular appearance (“W-” and ”V-view”).
Figure 5.
Electrostatic surfaces comparison of the superhelical grooves of the p115GHR, β-catenin and karyopherin-α.
Blue indicates positive charge and red negative charge at the level of 10 kT/e. The amino terminus of the molecule is at the top of the figure.
Figure 6.
Model of the overall fold of the full-length p115.
Surface and cartoon representations of a model of the full-length general vesicular transport factor p115 generated by manually fitting a coiled-coil of appropriate length to the C-termini of p115GHR in the crystallographic dimer. The different views of the p115 model closely resemble published electron micrographs of p115 and Uso1p.
Table 1.
Data collection and refinement statistics.