title: Tools for macromolecular model building and refinement into electron cryo-microscopy reconstructions.
abstract: The recent rapid development of single-particle electron cryo-microscopy (cryo-EM) now allows structures to be solved by this method at resolutions close to 3 Å. Here, a number of tools to facilitate the interpretation of EM reconstructions with stereochemically reasonable all-atom models are described. The BALBES database has been repurposed as a tool for identifying protein folds from density maps. Modifications to Coot, including new Jiggle Fit and morphing tools and improved handling of nucleic acids, enhance its functionality for interpreting EM maps. REFMAC has been modified for optimal fitting of atomic models into EM maps. As external structural information can enhance the reliability of the derived atomic models, stabilize refinement and reduce overfitting, ProSMART has been extended to generate interatomic distance restraints from nucleic acid reference structures, and a new tool, LIBG, has been developed to generate nucleic acid base-pair and parallel-plane restraints. Furthermore, restraint generation has been integrated with visualization and editing in Coot, and these restraints have been applied to both real-space refinement in Coot and reciprocal-space refinement in REFMAC.
authors.affiliation: MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, England.
authors.forename: Alan
authors.initials: A
authors.lastname: Brown
authors.affiliation: MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, England.
authors.forename: Fei
authors.initials: F
authors.lastname: Long
authors.affiliation: MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, England.
authors.forename: Robert A
authors.initials: RA
authors.lastname: Nicholls
authors.affiliation: MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, England.
authors.forename: Jaan
authors.initials: J
authors.lastname: Toots
authors.affiliation: MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, England.
authors.forename: Paul
authors.initials: P
authors.lastname: Emsley
authors.affiliation: MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, England.
authors.forename: Garib
authors.initials: G
authors.lastname: Murshudov
mesh_terms: D020285:Cryoelectron Microscopy / Q000379:methods*; D018360:Crystallography, X-Ray; D046911:Macromolecular Substances / Q000737:chemistry*; D008958:Models, Molecular
chemical_list: D046911:Macromolecular Substances
keywords: LIBG; electron cryo-microscopy reconstructions; model building; refinement