Atomic modeling of cryo-electron microscopy reconstructions--joint refinement of model and imaging parameters

J Struct Biol. 2013 Apr;182(1):10-21. doi: 10.1016/j.jsb.2013.01.003. Epub 2013 Jan 29.

Abstract

When refining the fit of component atomic structures into electron microscopic reconstructions, use of a resolution-dependent atomic density function makes it possible to jointly optimize the atomic model and imaging parameters of the microscope. Atomic density is calculated by one-dimensional Fourier transform of atomic form factors convoluted with a microscope envelope correction and a low-pass filter, allowing refinement of imaging parameters such as resolution, by optimizing the agreement of calculated and experimental maps. A similar approach allows refinement of atomic displacement parameters, providing indications of molecular flexibility even at low resolution. A modest improvement in atomic coordinates is possible following optimization of these additional parameters. Methods have been implemented in a Python program that can be used in stand-alone mode for rigid-group refinement, or embedded in other optimizers for flexible refinement with stereochemical restraints. The approach is demonstrated with refinements of virus and chaperonin structures at resolutions of 9 through 4.5 Å, representing regimes where rigid-group and fully flexible parameterizations are appropriate. Through comparisons to known crystal structures, flexible fitting by RSRef is shown to be an improvement relative to other methods and to generate models with all-atom rms accuracies of 1.5-2.5 Å at resolutions of 4.5-6 Å.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Archaeal Proteins / chemistry*
  • Archaeal Proteins / ultrastructure
  • Chaperonins / chemistry*
  • Chaperonins / ultrastructure
  • Cryoelectron Microscopy / methods*
  • Dependovirus / ultrastructure*
  • Fourier Analysis
  • Image Processing, Computer-Assisted
  • Immunoglobulin Fab Fragments / chemistry*
  • Immunoglobulin Fab Fragments / ultrastructure
  • Methanococcus / chemistry
  • Models, Molecular
  • Protein Structure, Tertiary

Substances

  • Archaeal Proteins
  • Immunoglobulin Fab Fragments
  • Chaperonins

Associated data

  • PDB/3J1Q
  • PDB/3J1S