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
Citations
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How Cryo-EM Became so Hot
TL;DR: Developing cryoelectron microscopy for the high-resolution structure determination of biomolecules in solution has broadened horizons for structural studies in molecular and cell biology.
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3.9 Å structure of the yeast Mec1-Ddc2 complex, a homolog of human ATR-ATRIP
Xuejuan Wang,Xuejuan Wang,Tingting Ran,Xuan Zhang,Jiyu Xin,Zhihui Zhang,Tengwei Wu,Weiwu Wang,Gang Cai,Gang Cai +9 more
TL;DR: The ataxia telangiectasia–mutated and Rad3-related (ATR) kinase is a master regulator of DNA damage response and replication stress in humans, but the mechanism of its activation remains unclear.
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Mechanisms of assembly and genome packaging in an RNA virus revealed by high-resolution cryo-EM
Emma L. Hesketh,Yulia Meshcheriakova,Kyle C. Dent,Pooja Saxena,Pooja Saxena,Rebecca F. Thompson,Joseph J.B. Cockburn,George P. Lomonossoff,Neil A. Ranson +8 more
TL;DR: Cryo-electron microscopy reconstructions for the wild-type virus and an empty virus-like particle are determined and built and reveal the C-terminal region of the small coat protein subunit, essential for virus assembly and which was missing from previously determined crystal structures, allowing a new model for genome encapsidation and capsid assembly to be developed.
Journal ArticleDOI
Building Proteins in a Day: Efficient 3D Molecular Structure Estimation with Electron Cryomicroscopy
TL;DR: A new Bayesian framework for cryo-EM structure estimation is presented that builds on modern stochastic optimization techniques to allow one to scale to very large datasets and introduces a novel Monte-Carlo technique that reduces the cost of evaluating the objective function during optimization by over five orders of magnitude.
Journal ArticleDOI
Atomic structure of a rhinovirus C, a virus species linked to severe childhood asthma
Yue Liu,Marchel G. Hill,Thomas Klose,Zhenguo Chen,Kelly Watters,Yury A. Bochkov,Wen Jiang,Ann C. Palmenberg,Michael G. Rossmann +8 more
TL;DR: Cryo-EM atomic structures of the full virion and native empty particle (NEP) of RV-C15a, a cell-adapted rhinovirus C species of picornaviruses, provide the basis for designing antiviral therapeutics and vaccines.
References
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