High-resolution cryo-EM: the nuts and bolts.
Cryogenic electron microscopy (cryo-EM) and single-particle analysis now enables the determination of high-resolution structures of macromolecular assemblies that have resisted X-ray crystallography and other approaches. Successful high-resolution structure determination by cryo-EM always depends on the quality of the protein sample. While structural heterogeneity remains a key challenge for cryo-EM, it also represents a rare opportunity to study the intrinsic conformational flexibility of macromolecular assemblies. Here, we review the key technological advancements that have made this 'resolution revolution' possible and give a concise overview of the technical challenges that needed to be overcome to allow high-resolution structure determination.
Department of Biochemistry and Molecular Biology, Biomedicine Discovery Institute, Monash University, Melbourne, Victoria, Australia; Australian Research Council Centre of Excellence in Advanced Molecular Imaging, Monash University, Melbourne, Victoria, Australia. Electronic address: dominika.elmlund@monash.edu.
Dominika
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Elmlund
Department of Biochemistry and Molecular Biology, Biomedicine Discovery Institute, Monash University, Melbourne, Victoria, Australia; Australian Research Council Centre of Excellence in Advanced Molecular Imaging, Monash University, Melbourne, Victoria, Australia.
Sarah N
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Le
Department of Biochemistry and Molecular Biology, Biomedicine Discovery Institute, Monash University, Melbourne, Victoria, Australia; Australian Research Council Centre of Excellence in Advanced Molecular Imaging, Monash University, Melbourne, Victoria, Australia. Electronic address: hanselmlund@monash.edu.
Hans
H
Elmlund
D016477:Artifacts; D020285:Cryoelectron Microscopy / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D009038:Motion; D059629:Signal-To-Noise Ratio