A new cryo-EM single-particle Ab initio reconstruction method visualizes secondary structure elements in an ATP-Fueled AAA+ motor

Summary, in English

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 660-kDa ATP-fueled AAA+ motor to 7.5 Å 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.

• Författare: Hans Elmlund; Joakim Lundqvist; Salam Al-Karadaghi; Mats Hansson; Hans Hebert; Martin Lindahl
• Avdelning/ar: Biokemi och Strukturbiologi
• Publiceringsår: 2008
• Språk: Engelska
• Sidor: 934-947
• Publikation/Tidskrift/Serie: Journal of Molecular Biology
• Volym: 375
• Issue: 4
• Länkar: http://dx.doi.org/10.1016/j.jmb.2007.11.028
• Dokumenttyp: Artikel i tidskrift
• Förlag: Elsevier
• Ämne: Biological Sciences
• Nyckelord: electron microscopy; cryo-EM; single particle; reconstruction; alignment
• Status: Published
• ISBN/ISSN/Övrigt: ISSN: 1089-8638