Martin Lindahl
Programmer
A new cryo-EM single-particle Ab initio reconstruction method visualizes secondary structure elements in an ATP-Fueled AAA+ motor
Author
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.
Department/s
- Biochemistry and Structural Biology
Publishing year
2008
Language
English
Pages
934-947
Publication/Series
Journal of Molecular Biology
Volume
375
Issue
4
Document type
Journal article
Publisher
Elsevier
Topic
- Biological Sciences
Keywords
- electron microscopy
- cryo-EM
- single particle
- reconstruction
- alignment
Status
Published
ISBN/ISSN/Other
- ISSN: 1089-8638