Extended Data Fig. 5: Coincident modes of DISP1 conformational flexibility revealed by 3D classification and 3D variability analysis (3DVA).
From: Dispatched uses Na+ flux to power release of lipid-modified Hedgehog
a, Overlay comparing front view of R and T conformations (khaki and salmon, respectively) from 3D classification. Major conformational changes are localized to the extracellular domains. b, Cut-away view showing the formation of a ‘kink’ in the back-side linker of the T conformation, with an accompanying shift of about one helix turn that breaks a hydrogen bond between linker residue H777 and the backbone carbonyl of K767. c, Extracellular view of the R and T conformations, highlighting the movement of secondary structure elements in ECD1 (> 5 Å) and ECD2 (~2 Å). The shift of ECD1 and the formation of the inter-ECD linker ‘kink’ appear intimately related. Numbers indicate distances (Å) between the Cα of F772 in R and T (marked by ▲ in b) and R382 and S898 (marked by ● and ○, respectively, in c). d, Comparison of the most extreme R and T conformations from 3D classification (right) and the two extreme ends of PC2, PC2+ (R-like) and PC2- (T-like), from 3DVA left) shows that a nearly identical mode of motion is captured by both techniques. e, Distributions of DISP1-A particles stratified by their 3D class along the PC2 axis of 3DVA, demonstrating colinearity between the 3DVA trajectory and the 3D subclasses ordered by apparent conformation. T1 and T3, and R1 and R2, respectively, are subclasses of T and R conformations resolved by 3D classification (Extended Data Fig. 3).
