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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Potent neutralization of hepatitis A virus reveals a receptor mimic mechanism and the receptor recognition site
Xiangxi Wang,Ling Zhu,Minghao Dang,Zhongyu Hu,Qiang Gao,Qiang Gao,Shuai Yuan,Yao Sun,Bo Zhang,Jingshan Ren,Abhay Kotecha,Thomas S. Walter,Junzhi Wang,Elizabeth E. Fry,David I. Stuart,Zihe Rao,Zihe Rao +16 more
TL;DR: High-resolution cryo-EM structures of HAV full and empty particles and of the complex of Hav with R10 Fab reveal the atomic details of antibody binding and point to a receptor recognition site at the pentamer interface, suggesting the use of a receptor mimic mechanism to neutralize virus infection.
Journal ArticleDOI
Structural basis for the inhibition of RecBCD by Gam and its synergistic antibacterial effect with quinolones.
Martin Wilkinson,Lucy Troman,Wan Ak Wan Nur Ismah,Yuriy Chaban,Matthew B. Avison,Mark S. Dillingham,Dale B. Wigley +6 more
TL;DR: High-resolution cryo-electron microscopy data explain the synthetic lethality observed between topoisomerase-induced DNA breaks and the RecBCD gene products, suggesting a new co-antibacterial strategy.
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Structural Basis of BRCC36 Function in DNA Repair and Immune Regulation.
Julius Rabl,Richard D. Bunker,Richard D. Bunker,Andreas D. Schenk,Andreas D. Schenk,Simone Cavadini,Simone Cavadini,M.E. Gill,M.E. Gill,W. Abdulrahman,W. Abdulrahman,A. Andres-Pons,A. Andres-Pons,Luijsterburg,Adel F. M. Ibrahim,Emma Branigan,Jacob D. Aguirre,Jacob D. Aguirre,Aimee H. Marceau,Claire Guérillon,Tewis Bouwmeester,Ulrich Hassiepen,Antoine H.F.M. Peters,Antoine H.F.M. Peters,Martin Renatus,L. Gelman,L. Gelman,Seth M. Rubin,Niels Mailand,H. van Attikum,Ronald T. Hay,Nicolas H. Thomä,Nicolas H. Thomä +32 more
TL;DR: This work explains modularity in the BRCC36 DUB family, with different adaptor subunits conferring diversified targeting and regulatory functions in the BRCA1-A and BRISC complexes.
Journal ArticleDOI
Performance of a direct detection camera for off-axis electron holography.
Shery L. Y. Chang,Shery L. Y. Chang,Christian Dwyer,Christian Dwyer,Juri Barthel,Chris Boothroyd,Rafal E. Dunin-Borkowski +6 more
TL;DR: It is demonstrated that the improved modulation transfer functions and detective quantum efficiencies of both modes of the DDC give rise to significant benefits over the conventional CCD cameras, specifically, a significant improvement in the visibility of the holographic fringes and a reduction of the statistical error in the phase of the reconstructed electron wave function.
Journal ArticleDOI
Insights into the Structure and Dynamics of Metal-Organic Frameworks via Transmission Electron Microscopy.
Xinyi Gong,Karthikeyan Gnanasekaran,Zhijie Chen,Lee Robison,Megan C. Wasson,Kyle C. Bentz,Seth M. Cohen,Omar K. Farha,Nathan C. Gianneschi,Nathan C. Gianneschi +9 more
TL;DR: A perspective on TEM studies of MOFs is provided and the utilization of state-of-the-art TEM technologies are highlighted to explore dynamic MOF processes and host-guest interactions.
References
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