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

Elmlund, Hans LU ; Lundqvist, Joakim LU ; Al-Karadaghi, Salam LU ; Hansson, Mats LU ; Hebert, Hans and Lindahl, Martin LU (2008) In Journal of Molecular Biology 375(4). p.934-947
Abstract
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... (More)
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. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
electron microscopy, cryo-EM, single particle, reconstruction, alignment
in
Journal of Molecular Biology
volume
375
issue
4
pages
934 - 947
publisher
Elsevier
external identifiers
  • wos:000253098000005
  • scopus:37449019135
ISSN
1089-8638
DOI
10.1016/j.jmb.2007.11.028
language
English
LU publication?
yes
id
add11e0b-479f-4984-a782-e039fb8da98f (old id 948914)
date added to LUP
2008-01-24 16:57:30
date last changed
2017-06-25 04:41:54
@article{add11e0b-479f-4984-a782-e039fb8da98f,
  abstract     = {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.},
  author       = {Elmlund, Hans and Lundqvist, Joakim and Al-Karadaghi, Salam and Hansson, Mats and Hebert, Hans and Lindahl, Martin},
  issn         = {1089-8638},
  keyword      = {electron microscopy,cryo-EM,single particle,reconstruction,alignment},
  language     = {eng},
  number       = {4},
  pages        = {934--947},
  publisher    = {Elsevier},
  series       = {Journal of Molecular Biology},
  title        = {A new cryo-EM single-particle Ab initio reconstruction method visualizes secondary structure elements in an ATP-Fueled AAA+ motor},
  url          = {http://dx.doi.org/10.1016/j.jmb.2007.11.028},
  volume       = {375},
  year         = {2008},
}