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The stereochemical quality of the final model was assessed with the programs MOLPROBITY [39] and PROCHECK [40], and agreement between the atomic model and X-ray data was verified with SFCHECK [41].
The FSC curve computed between the atomic model and the test map is shown in blue.
We added to Figure 2 the FSC curve computed between the atomic model and the test map.
A recent paper in eLife (Xu et al., 2014) almost certainly imposed an incorrect helical symmetry and this can be seen using filament images posted by Xu et al. A comparison between the atomic model proposed and the published three-dimensional reconstruction should have suggested that an incorrect solution was found.
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The crystal structure of the Escherichia coli 70S ribosome (Schuwirth et al, 2005) was fit into the cryo-EM density map using real-space refinement, a procedure which locally optimizes the cross-correlation between pieces of the atomic model and the density map (Gao et al, 2003).
Explicitly modelled solvent atoms contribute to the atomic model (F calc ).
(C ) Fourier shell correlation between the map generated from the atomic model (panel B, right) and the CTF corrected (panel B, center), and uncorrected (panel B, left) subtomogram averages.
The fitting is performed by iteratively deforming the initial structure along a set of low-frequency normal modes, which increases the cross-correlation coefficient (CCC) between the calculated electron density from the atomic model and the observed density from the EM map.
Therefore, an important difference between model building in crystallography and in EM is that the atomic model is not required to refine the EM map.
(C ) FSC between the segmented cryoEM map of one CARD and the map calculated from the atomic model (PDB: 3J6J, residues 1 97).
T.O. and F.W. generated the atomic model.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com