期刊
STRUCTURE
卷 18, 期 6, 页码 667-676出版社
CELL PRESS
DOI: 10.1016/j.str.2010.05.001
关键词
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资金
- German Federal Ministry of Education and Research (BMBF)
- European 3D Repertoire grant
- Ernst-Jung-Stiftung
- Fonda der Chemischen Industrie
- Novo Nordisk Foundation
- Agnes og Poul Friis' Foundation
- Fru Astrid Thaysens Legat for Laegevidenskabelig Grundforskning
- A.P. Moller Foundation for the Advancement of Medical Sciences
- Danish Council for Independent Research, Medical Sciences
- Danish Center for Scientific Computing (DCSC)
Single-particle electron cryomicroscopy is a powerful method for three-dimensional (3D) structure determination of macromolecular assemblies. Here we address the challenge of determining a 3D structure in the absence of reference models. The 3D structures are determined by alignment and weighted averaging of densities obtained by native cryo random conical tilt (RCT) reconstructions including consideration of missing data. Our weighted averaging scheme (wRCT) offers advantages for potentially heterogeneous 3D densities of low signal-to-noise ratios. Sets of aligned RCT structures can also be analyzed by multivariate statistical analysis (MSA) to provide insights into snapshots of the assemblies. The approach is used to compute 3D structures of the Escherichia coli 70S ribosome and the human U4/U6.U5 tri-snRNP under vitrified unstained cryo conditions, and to visualize by 3D MSA the L7/L12 stalk of the 70S ribosome and states of tri-snRNP. The approach thus combines de novo 3D structure determination with an analysis of compositional and conformational heterogeneity.
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