A new cryo-EM single-particle ab initio reconstruction method visualizes secondary structure elements in an ATP-fueled AAA+ motor
2008 (English)In: Journal of Molecular Biology, ISSN 0022-2836, E-ISSN 1089-8638, Vol. 375, no 4, 934-947 p.Article in journal (Refereed) Published
The generation of ab initio three-dimensional (3D) models is a bottleneck in the studies of large macromolecular assemblies by single-particle cryo-electron microscopy. We describe here a novel method, in which established methods for two-dimensional image processing are combined with newly developed programs for joint rotational 3D alignment of a large number of class averages (RAD) and calculation of 3D volumes from aligned projections (VolRec). We demonstrate the power of the method by reconstructing an similar to 660-kDa ATP-fueled AAA+ motor to 7.5 angstrom resolution, with secondary structure elements identified throughout the structure. We propose the method as a generally applicable automated strategy to obtain 3D reconstructions from unstained single particles imaged in vitreous ice.
Place, publisher, year, edition, pages
2008. Vol. 375, no 4, 934-947 p.
electron microscopy; cryo-EM; single particle; reconstruction; alignment
IdentifiersURN: urn:nbn:se:kth:diva-8111DOI: 10.1016/j.jmb.2007.11.028ISI: 000253098000005OAI: oai:DiVA.org:kth-8111DiVA: diva2:13343
QC 201007192008-03-182008-03-182010-10-01Bibliographically approved