A new cryo-EM single-particle Ab initio reconstruction method visualizes secondary structure elements in an ATP-Fueled AAA+ motor
(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:
https://lup.lub.lu.se/record/948914
- author
- Elmlund, Hans LU ; Lundqvist, Joakim LU ; Al-Karadaghi, Salam LU ; Hansson, Mats LU ; Hebert, Hans and Lindahl, Martin LU
- organization
- publishing date
- 2008
- 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
- pmid:18068723
- 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
- 2016-04-04 10:10:12
- date last changed
- 2025-04-04 14:49:54
@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}},
keywords = {{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}},
doi = {{10.1016/j.jmb.2007.11.028}},
volume = {{375}},
year = {{2008}},
}