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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Tubulin glycylation controls axonemal dynein activity, flagellar beat, and male fertility
Sudarshan Gadadhar,Sudarshan Gadadhar,Gonzalo Alvarez Viar,Jan N. Hansen,An Gong,Aleksandr Kostarev,Côme Ialy-Radio,Sophie Leboucher,Sophie Leboucher,Marjorie Whitfield,Ahmed Ziyyat,Aminata Touré,Luis Alvarez,Gaia Pigino,Carsten Janke,Carsten Janke +15 more
TL;DR: In this paper, the importance of microtubule glycylation for controlled flagellar beating, directional sperm swimming, and male fertility was revealed. But its functional and mechanistic roles remain unclear, and their perturbations have been linked to a growing number of human pathologies.
Journal ArticleDOI
Structural insights into Ca2+-activated long-range allosteric channel gating of RyR1
Risheng Wei,Xue Wang,Yan Zhang,Saptarshi Mukherjee,Lei Zhang,Qiang Chen,Xinrui Huang,Shan Jing,Congcong Liu,Shuang Li,Guangyu Wang,Yaofang Xu,Sujie Zhu,Alan J. Williams,Fei Sun,Chang-Cheng Yin +15 more
TL;DR: The characterization of a Ca2+-activated open-state RyR1 structure by cryo-electron microscopy provides structural insights into the molecular mechanisms of channel gating and regulation of RyRs and sheds light on structural basis for channel-gating and ion selectivity mechanisms for the six-transmembrane-helix cation channel family.
Journal ArticleDOI
Ionization and structural properties of mRNA lipid nanoparticles influence expression in intramuscular and intravascular administration.
Manuel J. Carrasco,Suman Alishetty,Mohamad-Gabriel Alameh,Hooda Said,Lacey Wright,Mikell Paige,Ousamah Younoss Soliman,Drew Weissman,Thomas E. Cleveland,Alexander Grishaev,Michael D. Buschmann +10 more
TL;DR: In this article, a method that can predict the pKa of lipid nanoparticles from the structure of the ionizable lipid was proposed, which can explain a wide range of delivery efficiencies in vitro and in vivo for intramuscular (IM) and intravascular (IV) administration of different ionizable lipids.
Journal ArticleDOI
Simultaneous Determination of Protein Structure and Dynamics Using Cryo-Electron Microscopy.
TL;DR: An integrative modeling approach is reported to simultaneously determine structure and dynamics of macromolecular systems from cryo-electron microscopy density maps and provide insights into the mechanisms by which the integral membrane receptor STRA6 interacts with retinol binding protein and translocatesretinol across the membrane.
Journal ArticleDOI
Cryo-EM structure of the bacteriophage T4 portal protein assembly at near-atomic resolution
Lei Sun,Xinzheng Zhang,Song Gao,Prashant Rao,Victor Padilla-Sanchez,Zhenguo Chen,Siyang Sun,Ye Xiang,Sriram Subramaniam,Venigalla B. Rao,Michael G. Rossmann +10 more
TL;DR: Comparison of the Myoviridae T4 portal structure with the known portal structures of φ29, SPP1 and P22, representing Podo- and Siphoviraceae, shows that the portal structure probably dates back to a time when self-replicating microorganisms were being established on Earth.
References
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