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Löffelholz, Ottilie von; Natchiar, S. Kundhavai; Djabeur, Nadia; Myasnikov, Alexander G.; Kratzat, Hanna; Menetret, Jean-Francois; Hazemann, Isabelle; Klaholz, Bruno P. (2017): Focused classification and refinement in high-resolution cryo-EM structural analysis of ribosome complexes. In: Current Opinion in Structural Biology, Vol. 46: pp. 140-148
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Cryo electron microscopy (cryo-EM) historically has had a strong impact on the structural and mechanistic analysis of protein synthesis by the prokaryotic and eukaryotic ribosomes. Vice versa, studying ribosomes has helped moving forwards many methodological aspects in single particle cryo-EM, at the level of automated data collection and image processing including advanced techniques for particle sorting to address structural and compositional heterogeneity. Here we review some of the latest ribosome structures, where cryo-EM allowed gaining unprecedented insights based on 3D structure sorting with focused classification and refinement methods helping to reach local resolution levels better than 3 angstrom. Such high-resolution features now enable the analysis of drug interactions with RNA and protein side-chains including even the visualization of chemical modifications of the ribosomal RNA. These advances represent a major breakthrough in structural biology and show the strong potential of cryo-EM beyond the ribosome field including for structure-based drug design.