Supplementary Figure 4: In silico sorting and resolution of the Api-RF1-70S complex.
From: An antimicrobial peptide that inhibits translation by trapping release factors on the ribosome
a, In silico sorting was performed with the FreAlign 9.11 software package (as described in Grigorieff, N., J. Struct. Biol. 157, 117-125 (2007)). Initial alignment of 116,212 particles was followed by 3D classification, resulting in six different classes. Class 1 (38,203 particles) was further refined, yielding a (b) final reconstruction consisting of 36,826 particles, with (c) an average resolution of 3.4 Å (based on the Fourier shell correlation (FSC) curve at FSC 0.143). d, Validation of the fit of molecular models to cryo-EM map for the Api137-RF1-70S complex. FSC curves calculated between the refined model and the final map (blue), with the self- and cross-validated correlations in orange and black, respectively. Information beyond 3.4 Å was not used during refinement and preserved for validation. (e) Side view and (f) transverse section of the cryo-EM map of Api137-RF1-70S complex colored according to local resolution as shown previously (Kucukelbir, A., Sigworth, F. J. & Tagare, H. D., Nat. Methods 11, 63-65 (2014)). g-h, Cryo-EM density for Api137 (g) colored according to local resolution and (h) shown as gray mesh with molecular model for residues 5-18.
