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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4.4 Å Resolution Cryo-EM structure of human mTOR Complex 1
Huirong Yang,Huirong Yang,Jia Wang,Mengjie Liu,Mengjie Liu,Xizi Chen,Xizi Chen,Min Huang,Dan Tan,Meng-Qiu Dong,Catherine C. L. Wong,Jiawei Wang,Yanhui Xu,Yanhui Xu,Hong-Wei Wang +14 more
TL;DR: The structure of human mTORC1 is determined at 4.4 Å resolution andStructural analysis indicates that FKBP12-Rapamycin may generate steric hindrance for substrate entry to the catalytic cavity of m TORC1.
ComponentDOI
Cryo-EM structure of the yeast respiratory supercomplex.
Sorbhi Rathore,Jens Berndtsson,Lorena Marín-Buera,Julian Conrad,Julian Conrad,Marta Carroni,Marta Carroni,Peter Brzezinski,Martin Ott +8 more
TL;DR: The overall architecture of the supercomplex is revealed, which deviates from the previously determined assemblies in mammals; the near-atomic structure of the yeast complex IV is obtained; and the protein-protein and protein-lipid interactions implicated in supercomplex formation are identified.
Journal ArticleDOI
Electron-event representation data enable efficient cryoEM file storage with full preservation of spatial and temporal resolution
Hui Guo,Erik Franken,Yuchen Deng,Samir Benlekbir,Garbi Singla Lezcano,Bart Janssen,Lingbo Yu,Zev A Ripstein,Yong Zi Tan,John L. Rubinstein +9 more
TL;DR: Electron-event representation is a new data format for cryoEM that preserves the full temporal and spatial resolution of movies from direct detector device cameras.
Journal ArticleDOI
Sub-2 Å Ewald curvature corrected structure of an AAV2 capsid variant
Yong Zi Tan,Sriram Aiyer,Mario Mietzsch,Joshua A. Hull,Robert McKenna,Joshua C Grieger,R. Jude Samulski,Timothy S. Baker,Mavis Agbandje-McKenna,Dmitry Lyumkis +9 more
TL;DR: It is demonstrated that Ewald sphere curvature correction, sub-Angstrom pixilation and per-particle CTF refinement can improve map quality and resolution and present the 1.86 Å cryo-EM structure of an adeno-associated virus serotype 2 variant.
Journal ArticleDOI
FEI's direct electron detector developments: Embarking on a revolution in cryo-TEM.
Maarten Kuijper,Gerald van Hoften,Bart Jozef Janssen,Rudolf Geurink,Sacha De Carlo,Matthijn R. J. Vos,Gijs van Duinen,Bart van Haeringen,Marc Storms +8 more
TL;DR: The signal formation in the sensor and its impact on the detection quantum efficiency (DQE) of the sensor is described and insights into the signal formation led to improved camera designs.
References
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