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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In Situ Structure of Neuronal C9orf72 Poly-GA Aggregates Reveals Proteasome Recruitment.
Qiang Guo,Carina Lehmer,Antonio Martinez-Sanchez,Till Rudack,Till Rudack,Florian Beck,Hannelore Hartmann,Manuela Pérez-Berlanga,Frédéric Frottin,Mark S. Hipp,F. Ulrich Hartl,Dieter Edbauer,Dieter Edbauer,Wolfgang Baumeister,Rubén Fernández-Busnadiego +14 more
TL;DR: Cryo-electron tomography is used to dissect the molecular architecture of protein aggregates within intact neurons at high resolution and finds that poly-GA aggregates consist of densely packed twisted ribbons that recruit numerous 26S proteasome complexes, while other macromolecules are largely excluded.
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The architecture of the mammalian respirasome
TL;DR: The structure of the respirasome provides information on the precise arrangements of the respiratory chain complexes in mitochondria as well as contributing to the oligomerization of CI and CIII.
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Structural Basis for Gating and Activation of RyR1.
Amedee des Georges,Amedee des Georges,Oliver B. Clarke,Ran Zalk,Qi Yuan,Kendall J. Condon,Kendall J. Condon,Robert A. Grassucci,Robert A. Grassucci,Wayne A. Hendrickson,Andrew R. Marks,Joachim Frank,Joachim Frank +12 more
TL;DR: Cryo-EM reconstructions of RyR1 in multiple functional states reveal the structural basis of channel gating and ligand-dependent activation, enabling analyses of conformational changes associated with gating.
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Structure of Mammalian Respiratory Supercomplex I1III2IV1
TL;DR: The cryo-electron microscopy structure of the major SC in porcine heart suggests a possible mechanism by which electrons are transferred from NADH to cytochrome c and provides a platform for future functional dissection of respiration.
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Architecture and conformational switch mechanism of the ryanodine receptor
TL;DR: The calcium-binding EF-hand domain is identified and it functions as a conformational switch allosterically gating the channel and represents a chimaera of voltage-gated sodium and pH-activated ion channels.
References
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