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Related Concept Videos

Cryo-electron Microscopy01:28

Cryo-electron Microscopy

Conventional electron microscopy (EM) involves dehydration, fixation, and staining of biological samples, which distorts the native state of biological molecules and results in several artifacts. Also, the high-energy electron beam damages the sample and makes it difficult to obtain high-resolution images. These issues can be addressed using cryo-EM, which uses frozen samples and gentler electron beams. The technique was developed by Jacques Dubochet, Joachim Frank, and Richard Henderson, for...

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Analyses of subnanometer resolution cryo-EM density maps.

Matthew L Baker1, Mariah R Baker, Corey F Hryc

  • 1National Center for Macromolecular Imaging, Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, Texas, USA.

Methods in Enzymology
|October 5, 2010
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Summary

Electron cryomicroscopy (cryo-EM) now achieves high resolutions for macromolecular assemblies. Computational tools help analyze cryo-EM density maps to build atomic models, aiding structural and functional insights.

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A Robust Single-Particle Cryo-Electron Microscopy (cryo-EM) Processing Workflow with cryoSPARC, RELION, and Scipion
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Area of Science:

  • Structural Biology
  • Biophysics
  • Computational Biology

Background:

  • Electron cryomicroscopy (cryo-EM) routinely achieves subnanometer resolutions for complex macromolecular assemblies.
  • Computational tools are essential for extracting structural and functional information from cryo-EM density maps.

Purpose of the Study:

  • To describe common computational tools for analyzing and modeling subnanometer resolution cryo-EM reconstructions.
  • To present a general protocol for analyzing cryo-EM density maps at subnanometer resolution.

Main Methods:

  • Utilizing computational analysis tools to interpret cryo-EM density maps.
  • Fitting atomic models into subnanometer resolution density maps.
  • Employing feature recognition for backbone and atomic model construction.

Main Results:

  • Subnanometer resolution allows isolation of subunits and identification of secondary structures.
  • Atomic models can be accurately fitted into density maps at resolutions beyond 5Å.
  • A protocol for analyzing 4.3Å resolution cryo-EM data of Mm-cpn is detailed.

Conclusions:

  • Computational tools are vital for deriving detailed structural information from cryo-EM data.
  • The described methods facilitate the construction of atomic models from subnanometer resolution cryo-EM maps.
  • Analysis of Mm-cpn at 4.3Å resolution demonstrates the utility of these computational approaches.