Challenges and opportunities in cryo-EM single-particle analysis

• Published in: The Journal of biological chemistry Vol. 294; no. 13; pp. 5181 - 5197
• Main Author: Lyumkis, Dmitry
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 29.03.2019; American Society for Biochemistry and Molecular Biology
• Subjects: Animals; atomic resolution; cryo-electron microscopy; Cryoelectron Microscopy - instrumentation; Cryoelectron Microscopy - methods; Equipment Design; Humans; Imaging, Three-Dimensional - instrumentation; Imaging, Three-Dimensional - methods; JBC Reviews; Protein Conformation; Protein Interaction Maps; protein structure; Proteins - chemistry; Proteins - metabolism; Proteins - ultrastructure; protein–protein interaction; single-particle analysis; structural biology; protein structure; protein–protein interaction; atomic resolution; structural biology; cryo-electron microscopy; single-particle analysis
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.REV118.005602

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.