Extended Data Fig. 6: Processing of the F-actin-R183W and F-actin-N111S datasets.
From: Molecular mechanisms of inorganic-phosphate release from the core and barbed end of actin filaments
a, g, Micrographs depicting R183W (a) and N111S (g) variants of F-actin frozen in vitreous ice, at, respectively, defocus values of −2.3 µm and −1.9 µm. The shown micrographs are example images from total datasets of 7,916 (R183W) and 9,516 (N111S) micrographs. The scale bars are 400 Å. b, h, Exemplary 2D-class averages of the R183W- (b) and N111S-F-actin (h) particles, computed through RELION. The box size is 267 \({\rm{\times }}\) 267 Å2. c, i, Image processing strategies that were employed to determine the actin-R183W (c) and actin-N111S (i) structures. All maps are shown in the same orientation. d, j, Angular distribution of the particles used to reconstruct the final cryo-EM maps of F-actin-R183W (d) and F-actin-N111S (j), shown along the filament axis (left) and orthogonal to the filament axis (right). e, k, Local-resolution estimations of the R183W- (e) and N111S-F-actin (k) density maps, calculated by RELION. The bar depicts local resolution in Å. f, l, Fourier-shell correlation plots for gold-standard refined masked (black), unmasked (blue) and high-resolution phase randomized (red) half-maps of F-actin-R183W (f) and F-actin-N111S particles (l). The FSC = 0.143 threshold is shown as a dashed line.