Validation, analysis and annotation of cryo‐EM structures. Issue 9 (2nd September 2021)

• Record Type: Journal Article
• Authors: Pintilie, Grigore; Chiu, Wah
• Is Part Of: Acta crystallographica. Volume 77:Issue 9(2021)
• Page Start: 1142
• Page End: 1152
• Publication Date: 2021-09-02
• Subjects: cryo‐EM -- validation -- Fourier shell correlation -- modeling -- annotation; X-ray crystallography -- Periodicals; Crystallography -- Periodicals; Molecular biology -- Periodicals; Molecular structure -- Periodicals; Biomolecules -- Structure -- Periodicals; Cytology -- Periodicals; Biomolecules -- Structure; Crystallography; Cytology; Molecular biology; Molecular structure; X-ray crystallography; Periodicals; 548
• DOI: 10.1107/S2059798321006069
• Languages: English
• ISSNs: 2059-7983

Abstract

Abstract : Methods for the validation of 3D maps from cryo‐EM are described and illustrated using an example map of bacteriophage P22. Methods for fitting, flexible fitting, refinement and annotation of models generated based on such maps are also explored. Abstract : The process of turning 2D micrographs into 3D atomic models of the imaged macromolecules has been under rapid development and scrutiny in the field of cryo‐EM. Here, some important methods for validation at several stages in this process are described. Firstly, how Fourier shell correlation of two independent maps and phase randomization beyond a certain frequency address the assessment of map resolution is reviewed. Techniques for local resolution estimation and map sharpening are also touched upon. The topic of validating models which are either built de novo or based on a known atomic structure fitted into a cryo‐EM map is then approached. Map–model comparison using Q ‐scores and Fourier shell correlation plots is used to assure the agreement of the model with the observed map density. The importance of annotating the model with B factors to account for the resolvability of individual atoms in the map is illustrated. Finally, the timely topic of detecting and validating water molecules and metal ions in maps that have surpassed ∼2 Å resolution is described.