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Martin Lindahl. Photo.

Martin Lindahl

Programmer

Martin Lindahl. Photo.

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

Author

  • Hans Elmlund
  • Joakim Lundqvist
  • Salam Al-Karadaghi
  • Mats Hansson
  • Hans Hebert
  • Martin Lindahl

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