Cryo-EM of macromolecular assemblies at near-atomic resolution. Academic Article uri icon

abstract

  • With single-particle electron cryomicroscopy (cryo-EM), it is possible to visualize large, macromolecular assemblies in near-native states. Although subnanometer resolutions have been routinely achieved for many specimens, state of the art cryo-EM has pushed to near-atomic (3.3-4.6 ) resolutions. At these resolutions, it is now possible to construct reliable atomic models directly from the cryo-EM density map. In this study, we describe our recently developed protocols for performing the three-dimensional reconstruction and modeling of Mm-cpn, a group II chaperonin, determined to 4.3 resolution. This protocol, utilizing the software tools EMAN, Gorgon and Coot, can be adapted for use with nearly all specimens imaged with cryo-EM that target beyond 5 resolution. Additionally, the feature recognition and computational modeling tools can be applied to any near-atomic resolution density maps, including those from X-ray crystallography.

published proceedings

  • Nat Protoc

altmetric score

  • 8

author list (cited authors)

  • Baker, M. L., Zhang, J., Ludtke, S. J., & Chiu, W.

citation count

  • 77

complete list of authors

  • Baker, Matthew L||Zhang, Junjie||Ludtke, Steven J||Chiu, Wah

publication date

  • September 2010