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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Near-atomic resolution for one state of f-actin.
Vitold E. Galkin,Albina Orlova,Matthijn R. J. Vos,Gunnar F. Schröder,Gunnar F. Schröder,Edward H. Egelman +5 more
TL;DR: The atomic model provides a framework for understanding why every buried residue in actin has been under intense selective pressure and insights into ATP-hydrolysis and filament dynamics.
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Invited Review Article: Methods for imaging weak-phase objects in electron microscopy
TL;DR: New activity in making an electron-optical equivalent of the familiar "phase-contrast" light microscope is based in part on the improved possibilities that are now available for device microfabrication, and a number of conceptually new phase-plate designs have been proposed, thus increasing the number of options that are available for development.
Journal ArticleDOI
Large-Scale Movements of IF3 and tRNA during Bacterial Translation Initiation.
Tanweer Hussain,José Luis Llácer,Brian T. Wimberly,Brian T. Wimberly,Jeffrey S. Kieft,Venki Ramakrishnan +5 more
TL;DR: CryoEM reconstructions of the complex of bacterial 30S subunit with initiator tRNA, mRNA, and IFs 1–3, representing different steps along the initiation pathway show that IF1 provides key anchoring points for IF2 and IF3, thereby enhancing their activities.
Journal ArticleDOI
Structure of mouse coronavirus spike protein complexed with receptor reveals mechanism for viral entry.
Jian Shang,Yushun Wan,Chang Liu,Boyd Yount,Kendra Gully,Yang Yang,Ashley Auerbach,Guiqing Peng,Ralph S. Baric,Fang Li +9 more
TL;DR: The cryo-EM structure of mouse hepatitis coronavirus spike complexed with mouse CEACAM1a is determined and a new role of receptor binding in MHV entry is revealed: in addition to its well-characterized role in viral attachment to host cells, receptor binding also induces the conformational change of the spike and hence the fusion of viral and host membranes.
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An atomic model of brome mosaic virus using direct electron detection and real-space optimization
Zhao Wang,Corey F. Hryc,Benjamin Bammes,Pavel V. Afonine,Joanita Jakana,Dong-Hua Chen,Xiangan Liu,Matthew L. Baker,C. Cheng Kao,Steven J. Ludtke,Michael F. Schmid,Paul D. Adams,Wah Chiu +12 more
TL;DR: A practical approach to obtain a rigorously validated atomic resolution electron cryo-microscopy structure using a newly implemented real-space optimization protocol for brome mosaic virus, an RNA virus.
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