Journal: Structure / Year: 2010 Title: An approach for de novo structure determination of dynamic molecular assemblies by electron cryomicroscopy. Authors: Bjoern Sander / Monika M Golas / Reinhard Lührmann / Holger Stark / Abstract: 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 ...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.
History
Deposition
May 10, 2010
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Header (metadata) release
Jun 11, 2010
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Map release
Jun 16, 2010
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Update
Jun 16, 2010
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Current status
Jun 16, 2010
Processing site: PDBe / Status: Released
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Structure visualization
Movie
Surface view with section colored by density value
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