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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From Electron Crystallography to Single Particle CryoEM (Nobel Lecture).
TL;DR: The development of electron microscopy from its beginnings to modern single particle cryo-EM is described by R. Henderson in his Nobel lecture and the first projection structure at 7 Å resolution of the purple membrane is shown.
Journal ArticleDOI
Development of a fast electromagnetic beam blanker for compressed sensing in scanning transmission electron microscopy
TL;DR: In this paper, an experimental setup based on an electromagnetic beam blanker placed in the condenser plane of a transmission electron microscopy (STEM) was proposed, where the blanker deflects the beam with a random pattern, while the scanning coils are moving the beam in the usual scan pattern.
ComponentDOI
Structure and architecture of immature and mature murine leukemia virus capsids.
K. Qu,Bärbel Glass,M. Dolezal,Florian K. M. Schur,Florian K. M. Schur,Brice Murciano,A. Rein,M. Rumlova,Tomáš Ruml,Hans-Georg Kräusslich,John A. G. Briggs,John A. G. Briggs +11 more
TL;DR: The dramatic architectural change on maturation indicates that extensive disassembly and reassembly are required for mature core growth in Murine leukemia virus, and suggests that wrapping of the genome in CA sheets may be sufficient to protect the MLV ribonucleoprotein during cell entry.
Journal ArticleDOI
Liquid-crystalline phase transitions in lipid droplets are related to cellular states and specific organelle association.
Julia Mahamid,Dimitry Tegunov,Andreas Maiser,Jan Arnold,Heinrich Leonhardt,Jürgen M. Plitzko,Wolfgang Baumeister +6 more
TL;DR: It is suggested that under mitotic arrest and starvation, relative CE levels increase, presumably due to the consumption of TAG metabolites for membrane synthesis and mitochondrial respiration, respectively, supported by direct visualization of LD–mitochondrial membrane contact sites.
Journal ArticleDOI
Bioactive Functionalized Monolayer Graphene for High-Resolution Cryo-Electron Microscopy.
Nan Liu,Jincan Zhang,Yanan Chen,Chuan Liu,Xing Zhang,Kui Xu,Jie Wen,Zhipu Luo,Shulin Chen,Peng Gao,Kaicheng Jia,Zhongfan Liu,Hailin Peng,Hong-Wei Wang +13 more
TL;DR: A new type of cryo-EM grids using bioactive-ligand functionalized single-crystalline monolayer graphene membranes as supporting films and functionalized graphene membrane (FGM) grids exhibit specific binding affinity to histidine (His)-tagged proteins and complexes are designed and produced.
References
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Crystal structure of the 20S proteasome from the archaeon T. acidophilum at 3.4 A resolution.
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Accurate determination of local defocus and specimen tilt in electron microscopy
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Prevention of overfitting in cryo-EM structure determination
Sjors H.W. Scheres,Shaoxia Chen +1 more
TL;DR: Analysis of simulated data with realistic signal-to-noise ratios indicates that the accuracy of the orientation determination is not affected by the exclusion of high-frequency terms, nor by the use of a model that is reconstructed from only half of the particles, as expected.