Electron counting and beam-induced motion correction enable near-atomic-resolution single-particle cryo-EM
Xueming Li,Paul Mooney,Shawn Q. Zheng,Shawn Q. Zheng,Christopher R. Booth,Michael B. Braunfeld,Michael B. Braunfeld,Sander Gubbens,David A. Agard,David A. Agard,Yifan Cheng +10 more
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TLDR
This approach determined a 3.3-Å-resolution structure of an ∼700-kDa protein with D7 symmetry, the Thermoplasma acidophilum 20S proteasome, showing clear side-chain density and greatly enhances image quality and data acquisition efficiency.Abstract:
In recent work with large high-symmetry viruses, single-particle electron cryomicroscopy (cryo-EM) has achieved the determination of near-atomic-resolution structures by allowing direct fitting of atomic models into experimental density maps. However, achieving this goal with smaller particles of lower symmetry remains challenging. Using a newly developed single electron-counting detector, we confirmed that electron beam-induced motion substantially degrades resolution, and we showed that the combination of rapid readout and nearly noiseless electron counting allow image blurring to be corrected to subpixel accuracy, restoring intrinsic image information to high resolution (Thon rings visible to ∼3 A). Using this approach, we determined a 3.3-A-resolution structure of an ∼700-kDa protein with D7 symmetry, the Thermoplasma acidophilum 20S proteasome, showing clear side-chain density. Our method greatly enhances image quality and data acquisition efficiency-key bottlenecks in applying near-atomic-resolution cryo-EM to a broad range of protein samples.read more
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Book ChapterDOI
Refinement of Atomic Structures Against cryo-EM Maps
TL;DR: This review describes some of the methods for atomic structure refinement (fitting) against medium/high-resolution single-particle cryo-EM reconstructed maps and several popular crystallographic methods, such as 2Fo-Fc maps, solvent flattening, and feature-enhanced maps (FEMs) for visualization and model (re)building.
Journal ArticleDOI
Limited dishevelled/Axin oligomerization determines efficiency of Wnt/β-catenin signal transduction.
Wei Kan,Michael D Enos,Elgin Korkmazhan,Stefan Muennich,Dong-Hua Chen,Melissa V. Gammons,Mansi Vasishtha,Mariann Bienz,Alexander R. Dunn,Georgios Skiniotis,William I. Weis +10 more
TL;DR: The relative affinities and stoichiometry of the DIX-DAX interaction provide a mechanism for efficient inhibition of β-catenin phosphorylation upon Axin recruitment to the Wnt receptor complex.
Journal ArticleDOI
DNA-Packing Portal and Capsid-Associated Tegument Complexes in the Tumor Herpesvirus KSHV.
Danyang Gong,Xinghong Dai,Jonathan Jih,Yun-Tao Liu,Yun-Tao Liu,Guo-Qiang Bi,Ren Sun,Z. Hong Zhou +7 more
TL;DR: Structural-based mutageneses confirm that a triplex deep binding groove for CATCs is a hotspot that holds promise for antiviral development and suggests a mechanism whereby the portal orchestrates procapsid formation and asymmetric long-range determination of CATC attachment during DNA packaging prior to pleomorphic tegumentation/envelopment.
Journal ArticleDOI
Cryo-EM structures of the ATP-bound Vps4 E233Q hexamer and its complex with Vta1 at near-atomic resolution.
Shan Sun,Lin Li,Fan Yang,Xiaojing Wang,Fenghui Fan,Mengyi Yang,Chunlai Chen,Xueming Li,Hong-Wei Wang,Sen-Fang Sui +9 more
TL;DR: The structures of the ATP-bound Vps4E233Q hexamer and its complex with the cofactor Vta1 (vps twenty associated 1) at resolutions of 3.9 and 4.2 Å are reported, determined by electron cryo-microscopy to provide important insights into the process of the ESCRT-III polymer disassembly by Vps 4.
Journal ArticleDOI
Toward a high-resolution structure of IP3R channel
TL;DR: Together these efforts provide the important structural information for this class of ion channels and serve as the basis for further studies aiming at understanding of the IP3R function.
References
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Journal ArticleDOI
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