Extended Data Figure 7. Three dimensional classification of APC/C^{Cdh1.Hsl1.Apc11-UbcH10}.

(a) The 3D classification started with 97,999 motion corrected particles, which were divided into five classes. The resultant classes were grouped into three categories: (i) 80.6% in the active ternary state with coactivator and substrate (Hsl1); (ii) 9.3% in the apo state and (iii) 10.1% in a hybrid state. Particles in the active ternary state (reconstructed to an overall resolution of 4.3 Å) were subjected to Cycle 2 classification with ten classes. UbcH10 density in the resultant classes is indicated with circles. (b) Classes with UbcH10 density in Cycle 2 classification are superimposed, showing variability of UbcH10. (c) Enlarged view of UbcH10 density. (d) A negative stain EM reconstruction of an APC/C^Cdh1.Hsl1 complex at ~25 Å resolution with a 1500-fold excess of UbcH10. The molecular surface is shown as a mesh representation and the coordinates of the APC/C^{Cdh1.Hsl1.UbcH10-Ub} were docked into the EM reconstruction. The UbcH10 coordinates fit new EM density proximal to Apc11.