Cryogenic electron microscopy and single-particle analysis.
About 20 years ago, the first three-dimensional (3D) reconstructions at subnanometer (<10-Å) resolution of an icosahedral virus assembly were obtained by cryogenic electron microscopy (cryo-EM) and single-particle analysis. Since then, thousands of structures have been determined to resolutions ranging from 30 Å to near atomic (<4 Å). Almost overnight, the recent development of direct electron detectors and the attendant improvement in analysis software have advanced the technology considerably. Near-atomic-resolution reconstructions can now be obtained, not only for megadalton macromolecular complexes or highly symmetrical assemblies but also for proteins of only a few hundred kilodaltons. We discuss the developments that led to this breakthrough in high-resolution structure determination by cryo-EM and point to challenges that lie ahead.
Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria 3800, Australia; email: hans.elmlund@monash.edu.
Dominika
D
Elmlund
Hans
H
Elmlund
D020285:Cryoelectron Microscopy / Q000295:instrumentation / Q000379:methods*; D005057:Eukaryotic Cells / Q000648:ultrastructure; D046911:Macromolecular Substances / Q000648:ultrastructure; D008958:Models, Molecular; D012270:Ribosomes / Q000648:ultrastructure; D012984:Software
D046911:Macromolecular Substances
3D reconstruction; ab initio; electron microscopy; image processing; near-atomic resolution; projection matching