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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Cryo-EM structure of the mammalian ATP synthase tetramer bound with inhibitory protein IF1
Jinke Gu,Laixing Zhang,Shuai Zong,Runyu Guo,Tianya Liu,Jingbo Yi,Peiyi Wang,Wei Zhuo,Maojun Yang,Maojun Yang +9 more
TL;DR: Porcine tetrameric ATP synthase is isolated and solved its structure at 6.2-angstrom resolution using a single-particle cryo-electron microscopy method, which is consistent with the ATP synthases tetramer adopting an inhibited state.
Journal ArticleDOI
Molecular dynamics-based refinement and validation for sub-5 Å cryo-electron microscopy maps
TL;DR: Two structure determination methods, based on the molecular dynamics flexible fitting (MDFF) paradigm, are presented that resolve sub-5 Å cryo-electron microscopy (EM) maps with either single structures or ensembles of such structures.
Journal ArticleDOI
Atomic model of the F420-reducing [NiFe] hydrogenase by electron cryo-microscopy using a direct electron detector
TL;DR: From the rigidity of the complex, it is concluded that catalysis is diffusion-limited and does not depend on protein flexibility or conformational changes.
Journal ArticleDOI
Rac1 GTPase activates the WAVE regulatory complex through two distinct binding sites
Baoyu Chen,Hui-Ting Chou,Chad A. Brautigam,Wenmin Xing,Sheng Yang,Lisa Henry,Lynda K. Doolittle,Thomas Walz,Michael K. Rosen +8 more
TL;DR: Cryo-electron microscopy data reveal that the WRC is activated by simultaneous engagement of two Rac1 molecules, suggesting a mechanism by which cells may sense the density of active Rac1 at membranes to precisely control actin assembly.
Journal ArticleDOI
Structure of the MacAB-TolC ABC-type tripartite multidrug efflux pump.
Anthony W. P. Fitzpatrick,Salomé Llabrés,Arthur Neuberger,James N. Blaza,Xiao Chen Bai,Ui Okada,Satoshi Murakami,Hendrik W. van Veen,Ulrich Zachariae,Sjors H.W. Scheres,Ben F. Luisi,Dijun Du +11 more
TL;DR: In this article, an electron-cryo-microscopy structure of the ABC-type tripartite assembly of Escherichia coli was presented at near-atomic resolution.
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