Supplementary Figure 3: Negative-stain reconstruction of dynactin.
From: Structural organization of the dynein–dynactin complex bound to microtubules
(a) A representative micrograph with individual particles circled in white, and reference-free 2D class averages on the right. (b) Class averages of dynactin with the flexible globular domain of p150Glued in a range of positions (indicated by yellow arrow). (c) On the left is a model of the 50 nm p150Glued coiled coil, with a filtered density representing the size of the CAP-Gly (PDB ID: 1TXQ). The globular domain in the class averages (right) is ∼20nm from the shoulder. The inset shows a focused class average of the globular domain, showing that the density is much larger than a CAP-Gly. (d) Surface rendering of the 24Å negative stain dynactin reconstruction shown from 4 different views (top panel), with corresponding forward projections of the reconstruction in the middle panel. Matching reference-free class averages are shown in the bottom panel. (e) FSC plot for the 3D reconstruction. The resolution using a 0.5 cutoff is ∼24Å. (f) Angular distribution of particles contributing to 3D reconstruction, with height of each cylinder related to the number of particles at that Euler angle.
