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J Biol Chem. 2019 Mar 29;294(13):5181-5197. doi: 10.1074/jbc.REV118.005602. Epub 2019 Feb 25.

Challenges and opportunities in cryo-EM single-particle analysis.

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From the Laboratory of Genetics and Helmsley Center for Genomic Medicine, The Salk Institute for Biological Studies, La Jolla, California 92037


Cryogenic electron microscopy (cryo-EM) enables structure determination of macromolecular objects and their assemblies. Although the techniques have been developing for nearly four decades, they have gained widespread attention in recent years due to technical advances on numerous fronts, enabling traditional microscopists to break into the world of molecular structural biology. Many samples can now be routinely analyzed at near-atomic resolution using standard imaging and image analysis techniques. However, numerous challenges to conventional workflows remain, and continued technical advances open entirely novel opportunities for discovery and exploration. Here, I will review some of the main methods surrounding cryo-EM with an emphasis specifically on single-particle analysis, and I will highlight challenges, open questions, and opportunities for methodology development.


atomic resolution; cryo-electron microscopy; protein structure; protein–protein interaction; single-particle analysis; structural biology

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