Electron cryomicroscopy of biological machines at subnanometer resolution

Structure. 2005 Mar;13(3):363-72. doi: 10.1016/j.str.2004.12.016.

Abstract

Advances in electron cryomicroscopy (cryo-EM) have made possible the structural determination of large biological machines in the resolution range of 6-9 angstroms. Rice dwarf virus and the acrosomal bundle represent two distinct types of machines amenable to cryo-EM investigations at subnanometer resolutions. However, calculating the density map is only the first step, and much analysis remains to extract structural insights and the mechanism of action in these machines. This paper will review the computational and visualization methodologies necessary for analysis (structure mining) of the computed cryo-EM maps of these machines. These steps include component segmentation, averaging based on local symmetry among components, density connectivity trace, incorporation of bioinformatics analysis, and fitting of high-resolution component data, if available. The consequences of these analyses can not only identify accurately some of the secondary structure elements of the molecular components in machines but also suggest structural mechanisms related to their biological functions.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.
  • Review

MeSH terms

  • Capsid Proteins / chemistry
  • Cryoelectron Microscopy*
  • Models, Molecular*
  • Multiprotein Complexes / chemistry*
  • Nanostructures / chemistry
  • Protein Structure, Secondary
  • Viruses / chemistry

Substances

  • Capsid Proteins
  • Multiprotein Complexes