eBIC for Industry is a recently inaugurated center offering professional cryo-EM services to the global pharmaceutical and biotech industry.
概述:
因其“发展了对溶液中生物分子获取高分辨率结构的冷冻电子显微术”，瑞典皇家科学院授予了 Jacques Dubochet，Joachim Frank 以及 Richard Henderson 2017 年度诺贝尔化学奖。2017 诺贝尔化学奖科学背景：冷冻电子显微术的发展
Gatan 的直接探测相机 (K3® 与 K2®) 背后的技术与创新，引领了冷冻电子显微术的进展。
什么是单颗粒冷冻电子显微术 (SINGLE-PARTICLE CRYO-EM)？
单颗粒冷冻电子显微术 (cryo-EM) 是一项逐步普及的技术，结构生物学家利用这项技术以原子级的分辨率进行结构解析。这项技术实现了对 X 射线晶体学的补充，因为它无需使用晶体样品即可显示结构细节。通过在玻璃态（非晶）冰中观察冷冻含水样品，可以保留天然状态的样品超微结构、缓冲液成分和配位体分布。冷冻电子显微术同时也是对核磁共振 (NMR) 结构研究的补充，因为它可研究大于 90 kDa 的样品。结构生物学家频繁使用冷冻电子显微术来研究病毒、小细胞器和生物大分子复合物，纯化蛋白以及超分子组装体或机器中的分子间相互作用。
在单颗粒冷冻电子显微术中，通过透射电子显微镜 (TEM) 记录每种样品的数千乃至数十万个相同但取向随机分布的微粒（分子）的高分辨率图像。然后使用图像分类算法对这些图像进行分组、对齐和平均，以区分三维分子的不同取向。对于优质样品，冷冻电子显微术能够以优于 1.5 Å的分辨率 解析分子结构；而仅仅几年之前，这样的分辨率水平还是令人不敢想象的。
冷冻电子显微术分辨率的迅速提升被称为“分辨率革命”，它是直接探测相机带来的直接结果。传统的电子显微术相机通过闪烁体将电子图像转换为光图像，并通过光纤面板将图像传输到 CCD 或 CMOS 图像传感器，并以模拟信号方式进行记录。结果经过这样的转换和记录后，会导致高分辨率细节丢失，这使得冷冻电子显微术一直无法发挥它的真正潜力。
如今，直接探测相机可以直接对电子图像进行观测，从而避免了传统相机信号转换步骤所造成的细节丢失。Gatan K3 相机是一款独具特色的直接探测相机，采用了超分辨率电子计数技术来记录图像。这项技术可以对单个的成像电子进行实时侦测和计数，同时排除模拟读出的噪声。正如 Gatan 的直接探测相机所示在 0.5 Nyquist 上的量子探测效率 (DQE)的优异表现。
分辨率的有效界限
Gatan 的直接探测相机，通常与 GIF Quantum LS 成像过滤系统结合使用，不断取得了刷新分辨率有效界限的突破性成果。许多前沿结果的发表，展示了运用这些产品，以及更优的数据采集和图像处理策略，对更小分子获取更高分辨率重构，例如β-半乳糖苷酶的 2.2 Å 结构。
上图显示了业已发表的单颗粒冷冻电子显微术获得的结构的翻遍率和分子量比较。在跨越了广泛分子量区间的分辨率前沿的结构中运用了 Gatan 的直接探测相机。
单颗粒冷冻电子显微术的优势
既然单颗粒冷冻电子显微术能够提供媲美 X 射线晶体学的结构分辨率，同时该技术具有许多独特的优势，增加了其对结构生物学家的吸引力：
| 功能 | 优势 |
|---|---|
| 在天然含水状态下检测结构 | 在生物相关环境中保存样品，包括样本浓度和缓冲液组分 |
| 支持研究较大的组装体 | 可用于表征大于 150 kDa 且包含多种亚单元，抑或非均质的或者极难结晶的分子 |
| 揭示原子级分辨率的结构 | 除了 α 螺旋和 β 褶板，还可观察不对称的侧链、氢键和水分子 |
| 控制化学环境 | 您可改变实验条件，以检测分子在不同功能状态下的行为 |
| 消除结晶步骤 | 避免耗时较长且不确定的制备步骤；缩短您的发表时间 |
Workflow for single-particle cryo-EM
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第 1 步：提纯 要使用单一微粒冷冻电子显微术研究分子，样品必须经过提纯并且结构必须完整，才能实现优质的 3D 重建。理想情况下，您需要将样品放在缓冲液中，以保持其生化活性。样品中的分子浓度应足够高，以便您能够在显微镜下观察分子，但是不能太高而使分子聚集。最后，应优化实验条件，以使待研究的分子达到统一的构象状态。 |
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第 2 步：骤降冷冻 每个样品都将被冷冻，以防止其在显微镜的真空里冻干。近乎瞬间冷冻可以防止形成水晶体，因为水晶体会破坏样品结构。 首先，将溶液中的少量样品放到 TEM 网格上，然后使用吸水纸轻轻吸掉多余的液体。然后将 TEM 网格插入液态乙烷或乙烷/丙烷混合液，迅速润湿样品，去除热量，然后产生非晶或玻璃态冰。图中显示了吸取液体和将样品浸没在液态冷却剂中之前的 Cryoplunge® 3 系统。 |
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第 3 步：转移到 TEM 中 冷冻之后，您可将样品转移到专用的 TEM 样品杆，使其保持液氮温度。为了防止样品受到污染，当您将样品装载到样品杆中时，使用冷冻工作台可以保护样品，然后从工作台转移到 TEM 的过程中，使用冷冻防护罩封住样品。如图所示的是插入到 TEM 中之前冷冻传输杆从工作台撤出的状态。 |
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第 4 步：样品成像 当暴露在电子环境中时，样品的结构完整性会遭到破坏。在高分辨率结构信息丢失之前，通常可以使用 10–30 e-/Å2 的总剂量。为了防止样品遭到破坏，可以在捕获图像之前，使用低剂量成像步骤导航至所需的区域并将电子束聚焦。 Gatan 的直接探测相机的电子计数和超分辨率模式带来的高 DQE，可帮助您获取脆弱生物样品的优异质量图像。这些图像具有高的信噪比，可帮助您在 3D 颗粒重构过程中辨别水分子、离子和配体结构。您还可以利用 Gatan 的直接探测相机的剂量分割功能，进一步提高图像质量，该功能以最高 75 帧/秒的速度保存全幅帧，以便随后用来校正样品移动和帮助最小化漂移的影响。 |
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第 5 步：分析和重构 成像之后，Gatan Microscopy Suite® 软件会帮助您进行分析，并以多种格式导出数据。Gatan 照相机获取的数据将导入各种第三方软件工具，用于进行 3D 重建和虚拟化，包括 EMAN、Frealign、Relion 及许多其他工具。图中显示了冷冻电子显微术 20S 蛋白酶体（2.8 Å 分辨率）的 3D 密度。 |
Sets a new standard for the efficient, high-throughput collection of low-dose, single-particle, cryo-EM datasets from Gatan’s cameras.
DigitalMicrograph，或称为 Gatan Microscopy Suite，驱动您的电子相机和其他附件以支持一系列重要应用，包括断层扫描、原位、谱学和衍射成像等。
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Scientists reach 2.2 Å using cryo-electron microscopy
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First 3D single-particle reconstruction of 20S Proteasome at 2.8 Å resolution
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First ~700 kDa protein structure with D7 symmetry identified at 3.3 Å resolution using cryo-EM
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First 3.4 Å TRPV1 structure solved by cryo-EM
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Potent neutralizing monoclonal antibodies directed to multiple epitopes on the SARS-CoV-2 spike
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Structure of the RNA-dependent RNA polymerase from COVID-19 virus
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Structure of the RNA-dependent RNA polymerase from COVID-19 virus
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Distinct conformational states of SARS-CoV-2 spike protein
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The Efficient Frontier of Resolution
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First 3D structure of human γ-secretase determined by cryo-EM at 4.5 Å resolution
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First 3.2 Å β-galactosidase structure solved by cryo-EM
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K2 Summit camera enables the study of heterogeneous particles by cryo-EM
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K2XP sensor takes the K2 Summit camera to the next level of performance
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Frozen-hydrated rotavirus double-layered particles
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Frozen-hydrated image of the Ndc80 complex decorated microtubules
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Cryo-EM images of T7 phage
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Guo, L.; Bi, W.; Wang, X.; Xu, W.; Yan, R.; Zhang, Y.; Zhao, K.; Li, Y.; Zhang, M.; Cai, X.; Jiang, S.; Xie, Y.; Zhou, Q.; Lu, L.; Dang, B.
Free fatty acid binding pocket in the locked structure of SARS-CoV-2 spike protein
Science
2020
Toelzer, C.; Gupta, K.; Yadav, S. K. N.: Borucu, U.; Davidson, A. D.; Williamson, M. K.; Shoemark, D. K.; Garzoni, F.; Staufer, O.; Milligan, R.; Capin, J.; Mulholland, A. J.; Spatz, J.; Fitzgerald, D.; Berger, I.; Schaffitzel, C.
An ultrapotent synthetic nanobody neutralizes SARS-CoV-2 by stabilizing inactive spike
Science
2020
Schoof, M.; Faust, B.; Saunders, R. A.; Sangwan, S.; Rezelj, V.; Hoppe, N.; Boone, M.; Billesbølle, C. B.; Puchades, C.; Azumaya, C. M.; Kratochvil, H. T.; Zimanyi, M.; Deshpande, I.; Liang, J.; Dickinson, S.; Nguyen, H. C.; Chio, C. M.; Merz, G. E.; Thompson, M. C.; Diwanji, D.; Schaefer, K.; Anand, A. A.; Dobzinski, N.; Zha, B. S.; Simoneau, C. R.; Leon, K.; White, K. M.; Chio, U. S.; Gupta, M.; Jin, M.; Li, F.; Liu, Y.; Zhang, K.; Bulkley, D.; Sun, M.; Smith, A. M.; Rizo, A. N.; Moss, F.; Brilot, A. F.; Pourmal, S.; Trenker, R.; Pospiech, T.; Gupta, S.; Barsi-Rhyne, B.; Belyy, V.; Barile-Hill, A. W.; Nock, S.; Liu, Y.; Krogan, N. J.; Ralston, C. Y.; Swaney, D. L.; García-Sastre, A.; Ott, M.; Vignuzzi, M.; QCRG Structural Biology Consortium; Walter, P.; Manglik, A.
Nature Communications
2020
Custódio, T. F.; Das, H.; Sheward, D. J.; Hanke, L.; Pazicky, S.; Pieprzyk, J.; Sorgenfrei, M.; Schroer, M. A.; Gruzinov, A. Y.; Jeffries, C. M.; Graewert, M. A.; Svergun, D. I.; Dobrev, N.; Remans, K.; Seeger, M. A.; McInerney, G. M.; Murrell, B.; Hällberg, B. M.; Löw, C.
The architecture of inactivated SARS-CoV-2 with postfusion spikes revealed by cryo-EM and cryo-ET
Structure
2020
Liu, C.; Mendonça, L.; Yang, Y.; Gao, Y.; Shen, C.; Liu, J.; Ni, T.; Ju, B.; Liu, C.; Tang, X.; Wei, J.; Ma, X.; Zhu, Y.; Liu, W.; Xu, S.; Liu, Y.; Yuan, J.; Wu, J.; Liu, Z.; Zhang, Z.; Liu, L.; Wang, P.; Zhang, P.
Cell Reports
2020
Zhou, T.; Teng, I. -T.; Olia, A. S.; Cerutti, G.; Gorman, J.; Nazzari, A.; Shi, W.; Tsybovsky, Y.; Wang, L.; Wang, S.; Zhang, B.; Zhang, Y.; Katsamba, P. S.; Petrova, Y.; Banach, B. B.; Fahad, A. S.; Liu, L.; Lopez Acevedo, S. N.; Madan, B.; de Souza, M. O.; Pan, X.; Wang, P.; Wolfe, J. R.; Yin, M.; Ho, D. D.; Phung, E.; DiPiazza, A.; Chang, L. A.; Abiona, O. M.; Corbett, K. S.; DeKosky, B. J.; Graham, B. S.; Mascola, J. R.; Misasi, J.; Ruckwardt, T.; Sullivan, N. J.; Shapiro, L.; Kwong, P. D.
De novo design of picomolar SARS-CoV-2 miniprotein inhibitors
Science
2020
Cao, L.; Goreshnik, I.; Coventry, B.; Case, J. B.; Miller, L.; Kozodoy, L.; Chen, R. R.; Carter, L.; Walls, A. C.; Park, Y. -J.; Strauch, E. -M.; Stewart, L.; Diamond, M. S.; Veesler, D.; Baker, D.
SARS-CoV-2 neutralizing antibody structures inform therapeutic strategies
Nature
2020
Barnes, C. O.; Jette, C. A.; Abernathy, M. E.; Dam, K. -M., A.; Esswein, S. R.; Gristick, H. B.; Malyutin, A. G.; Sharaf, N. G.; Huey-Tubman, K. E.; Lee, Y. E.; Robbiani, D. F.; Nussenzweig, M. C.; West Jr., A. P.; Bjorkman, P. J.
In situ structural analysis of SARS-CoV-2 spike reveals flexibility mediated by three hinges
Science
2020
Turonova, B.; Sikora, M.; Schurmann, C.; Hagen, W. J. H.; Welsch, S.; Blanc, F. E. C.; von Bulow, S.; Gecht, M.; Bagola, K.; Horner, C.; van Zandbergen, G.; Landry, J.; de Azevedo, N. T. D.; Mosalaganti, S.; Schwarz, A.; Covino, R.; Muhlebach, M. D.; Hummer, G.; Locker, J, K,; Beck, M.
Broad host range of SARS-CoV-2 and the molecular basis for SARS-CoV-2 binding to cat ACE2
Cell Discovery
2020
Wu, L.; Chen, Q.; Liu, K.; Wang, J.; Han, P.; Zhang, Y.; Hu, Y.; Meng, Y.; Pan, X.; Qiao, C.; Tian, S.; Du, P.; Song, H.; Shi, W.; Qi, J.; Wang, H. -W.; Yan, J.; Gao, G. F.; Wang, Q.
Structure-based design of prefusion-stabilized SARS-CoV-2 spikes
Science
2020
Hseih, C. -L.; Goldsmith, J. A.; Schaub, J. M.; Divenere, A. M.; Kuo, H. -C.; Javanmardi, K.; Le, K. C.; Wrapp, D.; Lee, A. G.; Liu, Y.; Chou, C. -W.; Byrne, P. O.; Hjorth, C. K.; Johnson, N. V.; Ludes-Meyers, J.; Nguyen, A. W.; Park, J.; Wang, N.; Amengor, D.; Lavinder, J. J.; Ippolito, G. C.; Maynard, J. A.; Finkelstein, I. J.; McLellan, J. S.
Structural basis for neutralization of SARS-CoV-2 and SARS-CoV by a potent therapeutic antibody
Science
2020
Lv, Z.; Deng, Y. -Q.; Ye, Q.; Cao, L.; Sun, C. -Y.; Fan, C.; Huang, W.; Sun, S.; Sun, Y.; Zhu, L.; Chen, Q.; Wang, N.; Nie, J.; Cui, Z.; Zhu, D.; Shaw, N.; Li, X. -F.; Li, Q.; Xie, L.; Wang, Y.; Rao, Z.; Qin, C. -F.; Wang, X.
Cell
2020
Chen, J.; Malone, B.; Llewellyn, E.; Grasso, M.; Shelton, P. M. M.; Olinares, P. D. B.; Maruthi, K.; Eng, E. T.; Vatandaslar, H.; Chait, B. T.; Kapoor, T. M.; Darst, S. A.; Campbell, E. A.
Receptor binding and priming of the spike protein of SARS-CoV-2 for membrane fusion
Nature
2020
Benton, D. J.; Wrobel, A. G.; Xu, P.; Roustan, C.; Martin, S. R.; Rosenthal, P. B.; Skehel, J. J.; Gamblin, S. J.
SARS-CoV-2 Nsp1 binds the ribosomal mRMA channel to inhibit translation
Nature Structural & Molecular Biology
2020
Schubert, K.; Karousis, E. D.; Jomaa, A.; Scaiola, A.; Echeverria, B.; Gurzeler, L. -A.; Leibundgut, M.; Thiel, V.; Mühlemann, O.; Ban, N.
Neutralization of SARS-CoV-2 by destruction of the prefusion spike
Cell Host & Microbe
2020
Huo, J.; Zhao, Y.; Ren, J.; Zhou, D.; Duyvesteyn, H. M. E.; Ginn, H. M.; Carrique, L.; Malinauskas, T.; Ruza, R. R.; Shah, P. N. M.; Tan, T. K.; Rijal, P.; Coombes, N.; Bewley, K. R.; Tree, J. A.; Radecke, J.; Paterson, N. G.; Supasa, P.; Mongkolsapaya, J.; Screaton, G. R.; Carroll, M.; Townsend, A.; Fry, E. E.; Owens, R. J.; Stuart, D. I.
An alpaca nanobody neutralizes SARS-CoV-2 by blocking receptor interaction
Nature Communications
2020
Hanke, L.; Perez, L. V.; Sheward, D. J.; Das, H.; Schulte, T.; Moliner-Morro, A.; Corcoran, M.; Achour, A.; Hedestam, G. B.; Hällberg, B. M.; Murrell, B.; McInerney, G. M.
Structural basis for translational shutdown and immune evasion by the Nsp1 protein of SARS-CoV-2
Science
2020
Thoms, M.; Buschauer, R.; Ameismeier, M.; Loepke, L.; Denk, T.; Hirschenberger, M.; Kratzat, H.; Hayn, M.; Mackens-Kiani, T.; Cheng, J.; Straub, J. H.; Sturzel, C. M.; Frohlich, T.; Berninghausen, O.; Becker, T.; Kirchhoff, F.; Sparrer, K. M.; Beckmann, R.
Molecular architecture of the SARS-CoV-2 virus
Cell
2020
Yao, H.; Song, Y.; Chen, Y.; Wu, N.; Xu, J.; Sun, C.; Zhang, J.; Weng, T.; Zhang, Z.; Wu, Z.; Cheng, L.; Shi, D.; Lu, X.; Lei, J.; Crispin, M.; Shi, Y.; Li, L.; Li, S.
Studies in humanized mice and convalescent humans yield a SARS-CoV-2 antibody cocktail
Science
2020
Hansen, J.; Baum, A.; Pascal, K. E.; Russo, V.; Giordano, S.; Wloga, E.; Fulton, B. O.; Yan, Y.; Koon, K.; Patel, K.; Chung, K. M.; ermann, A.; Ullman, E.; Cruz, J.; Rafique, A.; Huang, T.; Fairhurst, J.; Libertiny, C.; Malbec, M.; Lee, W. -Y.; Welsh, R.; Farr, G.; Pennington, S.; Deshpande, D.; Cheng, J.; Watty, A.; Bouffard, P.; Babb, R.; Levenkova, N.; Chen, C.; Zhang, B.; Hernandez, A. R.; Saotome, K.; Zhou, Y.; Franklin, M.; Sivapalasingam, S.; Lye, D. C.; Weston, S.; Lopgue, J.; Haupt, R.; Frieman, M.; Chen., G.; Olson, W.; Murphy, A. J.; Stahl, N.; Yancopoulos, G. D.; Kyratsous, C. A.
Cell
2020
Barnes, C. O.; West Jr., A. P.; Huey-Tubman, K. E.; Hoffmann, M. A. G.; Sharaf, N. G.; Hoffman, P. R.; Koranda, N.; Gristick, H. B.; Gaebler, C.; Muecksch, F.; Lorenzi, J. C. C.; Finkin, S.; Hägglöf, T.; Hurley, A.; Millard, K. G.; Weisblum, Y.; Schmidt, F.; Hatziioannou, T.; Bieniasz, P. D.; Caskey, M.; Robbiani, D. F.; Nussenzweig, M. C.; Bjorkman. P. J.
A neutralizing human antibody binds to the N-terminal domain of the spike protein of SARS-CoV-2
Science
2020
Chi, X.; Yan, R.; Zhang, G.; Zhang, Z.; Hao, M.; Zhang, Z.; Fan, P.; Dong, X.; Yang, Y.; Chen, Z.; Guo, Y.; Zhang, J.; Li, Y.; Song, X.; Cheng, Y.; Xia, L.; Fu, L.; Hou, L.; Xu, J.; Yu, X.; Li, J.; Zhou, Q.; Chen, W.
Structural basis for the neutralization of SARS-CoV-2 by an antibody from a convalescent patient
Nature Structural & Molecular Biology
2020
Zhou, D.; Duyvesteyn, H. M. E.; Chen, C. -P.; Huang, C. -G.; Chen, T. -H.; Shih, S. -R.; Lin, Y. -C.; Cheng, C. -Y.; Cheng, S. -H.; Huang, Y. -C.; Lin, T. -Y.; Ma, C.; Huo, J.; Carrique, L.; Malinauskas, T.; Ruza, R. R.; Shah, P. N. M.; Tan, T. K.; Rijal, P.; Donat, R. F.; Godwin, K.; Buttigieg, K. R.; Tree, J. A.; Radecke, J.; Paterson, N. G.; Supasa, P.; Mongkolsapaya, J.; Screaton, G. R.; Carroll, M. W.; Gilbert-Jaramillo, J.; Knight, M. L.; James, W.; Owens, R. J.; Naismith, J. H.; Townsend, A. R.; Fry, E. E.; Zhao, Y.; Ren, J.; Stuart, D. I.; Huang, K. -Y. A.
A thermostable, closed SARS-CoV-2 spike protein trimer
Nature Structural & Molecular Biology
2020
Xiong, X.; Qu, K.; Ciazynska, K. A.; Hosmillo, M.; Carter, A. P.; Ebrahimi, S.; Ke, Z.; Scheres, S. H. W.; Bergamaschi, L.; Grice, G. L.; Zhang, Y.; The CITIID-NIHR COVID-19 BioResource Collaboration; Nathan, J. A.; Baker, S.; James, L. C.; Baxendale, H. E.; Goodfellow, I.; Doffinger, R.; Briggs, J. A. G.
Structural basis for RNA replication by the SARS-CoV-2 polymerase
Cell
2020
Wang, Q.; Wu2, J.; Wang, H.; Gao, Y.; Liu, Q.; Mu, A.; Ji, W.; Yan, L.; Zhu, Y.; Zhu, C.; Fang, X.; Yang, X.; Huang, Y.; Gao, H.; Liu, F.; Ge, J.; Sun, Q.; Yang, X.; Xu, W.; Liu, Z.; Yang, H.; Lou, Z.; Jiang, B.; Guddat, L. W.; Gong, P.; Rao, Z.
Controlling the SARS-CoV-2 spike glycoprotein conformation
Nature Structural & Molecular Biology
2020
Henderson, R.; Edwards, R. J.; Mansouri, K.; Janowska, K.; Stalls, V.; Gobeil, S. M. C.; Kopp, M.; Li, D.; Parks, R.; Hsu, A. L.; Borgnia, M. J.; Haynes, B. F.; Acharya, P.
Cryo-EM analysis of the post-fusion structure of the SARS-CoV spike glycoprotein
Nature Communications
2020
Fan, X.; Cao, D.; Kong, L.; Zhang, X.
Potently neutralizing and protective human antibodies against SARS-CoV-2
Nature
2020
Zost, S. J.; Gilchuk, P.; Case, J. B.; Binshtein, E.; Chen, R. E.; Nkolola, J. P.; Schäfer, A.; Reidy, J. X.; Trivette, A.; Nargi, R. S.; Sutton, R. E.; Suryadevara, N.; Martinez, D. R.; Williamson, L. E.; Chen, E. C.; Jones, T.; Day, S.; Myers, L.; Hassan, A. O.; Kafai, N. M.; Winkler, E. S.; Fox, J. M.; Shrihari, S.; Mueller, B. K.; Meiler, J.; Chandrashekar, A.; Mercado, N. B.; Steinhardt, J. J.; Ren, K.; Loo, Y. -M.; Kallewaard, N. L.; McCune, B. T.; Keeler, S. P.; Holtzman, M. J.; Barouch, D. H.; Gralinski, L. E.; Baric, R. S.; Thackray, L. B.; Diamond, M. S.; Carnahan, R. H.; Crowe Jr, J. E.
Cell
2020
Cao, Y.; Su, B.; Guo, X.; Sun, W.; Deng, Y.; Bao, L.; Zhu, Q.; Zhang, X.; Zheng, Y.; Geng, C.; Chai, X.; He, R.; Li, X.; Lv, Q.; Zhu, H.; Deng, W.; Xu, Y.; Wang, Y.; Qiao, L.; Tan, Y.; Song, L.; Wang, G.; Du, X.; Gao, N.; Liu, J.; Xiao, J.; Su, X. -D.; Du, Z.; Feng, Y.; Qin, C.; Qin, C.; Jin, R.; Xie, X. S.
Structural basis for inhibition of the RNA-dependent RNA polymerase from SARS-CoV-2 by remdesivir
Science
2020
Yin, W.; Mao, C.; Luan, X.; Shen, D. -D.; Shen, Q.; Su, H.; Wang, X.; Zhou, F.; Zhao, W.; Gao, M.; Chang, S.; Xie, Y. -C.; Tian, G.; Jiang, H. -W.; Tao, S. -C.; Shen, J.; Jiang, Y.; Jiang, H.; X.u, Y.; Zhang, S.; Xu, H. E.
eLife
2020
Langer, L. M.; Gat, Y.; Bonneau, F.; Conti, E.
Structure of replicating SARS-CoV-2 polymerase
Nature
2020
Hillen, H. S.; Kokic, G.; Farnung, L.; Dienemann, C.; Tegunov, D.; Cramer, P.
Structure, function, and antigenicity of the SARS-CoV-2 spike glycoprotein
Cell
2020
Walls, A. C.; Park, Y. -J.; Tortorici, M. A.; Wall, A.; McGuire, A. T.; Veesler, D.
Structural basis for the recognition of the SARS-CoV-2 by full-length human ACE2
Science
2020
Yan, R.; Zhang, Y.; Li, Y.; Xia, L.; Guo, Y.; Zhou, Q.
Structure, function and antigenicity of the SARS-CoV-2 spike glycoprotein
bioRxiv
2020
Walls, A. C.; Park, Y. -J.; Tortorici, M. A.; Wall, A.; McGuire, A. T.; Veesler, D
Communications Biology
2019
Celia, H.; Botos, I.; Ni, X.; Fox, T.; De Val, N.; Lloubes, R.; Jiang, J.; Buchanan, S. K.
Improved applicability and robustness of fast cryo-electron tomography data acquisition
Journal of Structural Biology
2019
Eisenstein, F.; Danev, R.; Pilhofer, M.
Surpassing the physical Nyquist limit to produce super-resolution cryo-EM reconstructions
bioRxiv
2019
Feathers, J. R.; Spoth, K. A.; Fromme, J. C.
Single particle cryo-EM reconstruction of 52 kDa streptavidin at 3.2 Angstrom resolution
Nature Communications
2019
Fan, X.; Wang, J.; Zhang, X.; Yang, Z.; Zhang, J. -C.; Zhao, L.; Peng, H. -L.; Lei, J.; Wang, H. -W.
Fusion-dependent formation of lipid nanoparticles containing macromolecular payloads
Nanoscale
2019
Kulkarni, J. A.; Witzigmann, D.; Leung, J.; van der Meel, R.; Zaifman, J.; Darjuan, M. M.; Grisch-Chan, H. M.; Thöny, B.; Tam, Y. Y. C.; Cullis, P. R.
ACS Omega
2019
Putti, M.; Stassen, O. M. J. A.; Schotman, M. J. G.; Sahlgren, C. M.; Dankers, P. Y. W.
A 3.8 Å resolution cryo-EM structure of a small protein bound to an imaging scaffold
Nature Communications
2019
Liu, Y.; Huynh, D. T.; Yeates, T. O.
In situ structures of polar and lateral flagella revealed by cryo-electron tomography
Journal of Bacteriology
2019
Zhu, S.; Schniederberend, M.; Zhitnitsky, D.; Jain, R.; Galán, J. E.; Kazmierczak, B. I.; Liu, J.
Refinement and analysis of the mature Zika virus cryo-EM structure at 3.1 Å resolution
Structure
2018
Sevvana, M.; Long, F.; Miller, A. S.; Klose, T.; Buda, G.; Sun, L.; Kuhn, R. J.; Rossmann, M. G.
Cryo-EM structure of 5-HT3A receptor in its resting conformation
Nature Communications
2018
Basak, S.; Gicheru, Y.; Samanta, A.; Molugu, S. K.; Huang, W.; la de Fuente, M.; Hughes, T.; Taylor, D. J.; Nieman, M. T.; Moiseenkova-Bell, V.; Chakrapani, S.
Structural mechanisms of centromeric nucleosome recognition by the kinetochore protein CENP-N
Science
2017
Chittori, S.; Hong, J.; Saunders, H.; Feng, H.; Ghirlando, R.; Kelly, A. E.; Bai, Y.
Cell
2017
Guo, T. W.; Bartesaghi, A.; Yang, H.; Falconieri, V.; Rao, P.; Merk, A.; Eng, E. T.; Raczkowski, A. M.; Fox, T.; Earl, L. A.; Patel, D. J.; Subramaniam, S.
Breaking cryo-EM resolution barriers to facilitate drug discovery
Cell
2017
Merk, A.; Bartesaghi, A.; Banerjee, S.; Falconieri,V.; Rao, P.; Davis, M.; Pragani, R.; Boxer, M.; Earl, L. A.; Milne, J. L. S.; Subramaniam, S.
Rules of engagement between alpha v beta 6 integrin and the foot-and-mouth disease virus
Nature Communications
2017
Kotecha, A.; Wang, Q.; Dong, X.; Ilca, S.; Ondiviela, M.; Zihe, R.; Seago, J.; Charleston, B.; Fry, E.; Abrescia, N.; Springer, T.; Huiskonen, J.; Stuart, D.
Cryo-EM structure of a separase-securin complex at near-atomic resolution
Nature Structural & Molecular Biology
2017
Boland, A.; Martin, T. G.; Ziguo, Z.; Yang, J.; Bai, X. C.; Chang, L.; Scheres, S. H. W.; Barford, D.
Atomic structure of the cystic fibrosis transmembrane conductance regulator
Cell
2016
Zhang, Z.; Chen, J.
PLOS ONE
2016
Zhao, H.; Speir, J. A.; Matsui, T.; Lin, Z.; Liang, L.; Lynn, A. Y.; Varnado, B.; Weiss, T. M.; Tang, L.
2.3 Å resolution cryo-EM structure of human p97 and mechanism of allosteric inhibition
Science
2016
Banerjee, S.; Bartesaghi, A.; Merk, A.; Rao, P.; Bulfer, S. L.; Yan, Y.; Green, N.; Mroczkowski, B.; Neitz, R. J.; Wipf, P.; Falconieri, V.; Deshaies, R. J.; Milne, J. L.; Huryn, D.; Arkin, M.; Subramaniam, S.
Structure of transcribing mammalian RNA polymerase II
Nature
2016
Bernecky, C.; Herzog, F.; Baumeister, W.; Plitzko, J. M.; Cramer, P.
A mechanism of viral immune evasion revealed by cryo-EM analysis of the TAP transporter
Nature
2016
Oldham, M. L., Hite, R. K.; Steffen, A. M.; Damko, E.; Li, Z.; Walz, T.; Chen, J.
Cryo-electron microscopy structure of the TRPV2 ion channel
Nature Structural & Molecular Biology
2016
Zubcevic, L.; Herzik, M. A.; Chung, B. C.; Liu, Z.; Lander, G. C.; Lee, S. -L.
Structures of a CRISPR-Cas9 R-loop complex primed for DNA cleavage
Science
2016
Jiang, F.; Taylor, D. W.; Chen, J. S.; Kornfeld, J. E.; Zhou, K.; Thompson, A. J.; Nogales, E.; Doudna, J. A.
Five of five VHHs neutralizing poliovirus bind the receptor-binding site
Journal of Virology
2016
Strauss, M.; Schotte, L.; Thys, B.; Filman, D. J.; Hogle, J. M.
Structure of Ljungan virus provides insight into genome packaging of this picornavirus
Nature Communications
2015
Zhu, L.; Wang, X.; Ren, J.; Porta, C.; Wenham, H.; Ekström, J. -O; Panjwani, A.; Knowles, N. J.; Kotecha, A.; Siebert, C. A.; Lindberg, A. M.; Fry, E. E.; Rao, Z.; Tuthill, T. J.; Stuart, D. I.
Dynamical features of the plasmodium falciparum ribosome during translation
Nucleic Acids Research
2015
Sun, M.; Li, W.; Blomqvist, K.; Das, S.; Hashem, Y.; Dvorin, J. D.; Frank, F.
Structure of a yeast spliceosome at 3.6-angstrom resolution
Science
2015
Yan, C.; Hang, J.; Wan, R.; Huang, M.; Wong, C. C. L.; Shi, Y.
An atomic structure of human c-secretase
Nature
2015
Bai, X. C.; Yan, C.; Yang, G.; Lu, P.; Ma, D.; Sun, L.; Zhou, R.; Scheres, S. H.; Shi, Y.
Mechanistic origin of microtubule dynamic instability and its modulation by EB proteins
Cell
2015
Zhang, R.; Alushin, G. M.; Brown, A.; Nogales, E.
In situ structural analysis of golgi intracisternal protein arrays
Proceedings of the National Academy of Sciences
2015
Engel, B. D.; Schaffer, M.; Albert, S.; Asano, S.; Plitzko, J. M.; Baumeister, W.
Structure of the L-protein of vesicular stomatitis virus from electron cryomicroscopy
Cell
2015
Liang, B.; Li, Z.; Jenni, S.; Rameh, A. A.; Morin, B. M.; Grant, T.; Grigorieff, N.; Harrison, S. C.; Whelan, S. P. J.
eLife
2015
Grant, T.; Grigorieff, N.
eLife
2015
Grant, T.; Grigorieff, N.
Electron cryomicroscopy observation of rotational states in a eukaryotic V-ATPase
Nature
2015
Zhao, J.; Benlekbir, S.; Rubinstein, J. L.
Structural basis of human γ-secretase assembly
Proceedings of the National Academy of Sciences
2015
Sun, L.; Zhao, L.; Yang, G.; Yan, C.; Zhou, R.; Zhou, X.; Xie, T.; Zhao, Y.; Wu, S.; Li, X.; Shi, Y.
Structures of the CRISPR-Cmr complex reveal mode of RNA target positioning
Science
2015
Taylor, D. W.; Zhu, Y.; Staals, R. H. J.; Kornfeld, J. E.; Shinkai, A.; van der Oost, J.; Nogales, E.; Doudna, J. A.
Structure of the TRPA1 ion channel suggests regulatory mechanisms
Nature
2015
Paulsen, C. E.; Armache, J.; Gao, Y.; Cheng, Y.; Julius, D.
Cryo-EM structure of influenza virus RNA polymerase complex at 4.3 Å resolution
Molecular Cell
2015
Chang, S.; Sun, D.; Liang, H.; Wang, J.; Li, J.; Guo, L.; Wang, X.; Guan, C.; Boruah, B. M.; Yuan, L.; Feng, F.; Yang, M.; Wang, L.; Wang, Y.; Wojdyla, J.; Li, L.; Wang, J.; Wang, M.; Cheng, G.; Wang, H. -W.; Liu, Y.
Nature Communications
2015
Fibriansah, G.; Tan, J. L.; Smith, S. A.; de Alwis, R.; Ng, T. S.; Kostyuchenko, V. A.; Jadi, R. S.; Kukkaro, P.; de Silva, A. M.; Crowe, J. E.; Lok, S. M.
Conformational changes leading to T7 DNA delivery upon interaction with the bacterial receptor
Journal of Biological Chemistry
2015
González-García, V. A.; Pulido-Cid, M.; Garcia-Doval, C.; Bocanegra, R.; van Raaij, M. J.; Martín-Benito, J.; Cuervo, A.; Carrascosa, J. L.
Mechanistic insights into the recycling machine of the SNARE complex
Nature
2015
Zhao, M.; Wu, S.; Zhou, Q.; Vivona, S.; Cipriano, D. J.; Cheng, Y.; Brunger, A. T.
Architecture of the RNA polymerase II-Mediator core transcription initiation complex
Nature
2015
Plaschka, C.; Lariviere, L.; Wenzeck, L.; Hemann, M.; Tegunov, D.; Petrotchenko, E. V.; Borchers, C. H.; Baumeister, W.; Herzog, F.; Villa, E.; Cramer, P.
Proceedings of the National Academy of Sciences
2015
Dai, X.; Gong, D.; Xiao, Y.; Wu, T. T.; Sun, R.; Zhou, Z. H.
A molecular census of 26S proteasomes in intact neurons
Science
2015
Asano, S.; Fukuda, Y.; Beck, F.; Aufderheide, A.; Förster, F.; Danev, R.; Baumeister, W.
Journal of Virology
2015
Strauss, M.; Filman, D. J.; Belnap, D. M.; Cheng, N.; Noel, R. T.; Hogle, J. M.
Native architecture of the Chlamydomonas chloroplast revealed by in situ cryo-electron tomography
eLife
2015
Engel, B. D.; Schaffer, M.; Cuellar, L. K.; Villa, E.; Plitzko, J. M.; Baumeister, W.
Rqc2p and the 60S ribosome mediate mRNA-independent elongation of nascent chains
Science
2015
Shen, P. S.; Park, J.; Qin, Y.; Li, X.; Parsawar, K.; Larson, M. H.; Cox, J.; Cheng, Y.; Lambowitz, A. M.; Weissman, J. S.; Brandman, O.; Frost, A.
Seeing tobacco mosaic virus through direct electron detectors
Journal of Structural Biology
2014
Fromm, S. A.; Bharat, T. A.; Jakobi, A. J.; Hagen, W. J.; Sachse, C.
Architecture of the ring formed by the tubulin homologue FtsZ in bacterial cell division
eLife
2014
Szwedziak, P.; Wang, Q.; Bharat, T.A.; Tsim, M.; Löwe, J.
eLife
2014
Shang, Z.; Zhou, K.; Xu, C.; Csencsits, R.; Cochran, J. C., Sindelar, C. V.
Allosteric communication in the dynein motor domain
Cell
2014
Bhabha, G.; Cheng, H.; Zhang, N.; Moeller, A.; Liao, M.; Speir, J. A.; Cheng, Y.; Vale, R. D.
Organization of capsid-associated tegument components in Kaposi's sarcoma-associated herpesvirus
Journal of Virology
2014
Dai, X.; Gong, D.; Wu, T. T.; Sun, R.; Zhou, Z. H.
Near-atomic resolution reconstructions using a mid-range electron microscope operated at 200 kV
Journal of Structural Biology
2014
Campbell, M. G.; Kearney, B. M.; Cheng, A.; Potter, C. S.; Potter, C. S.; Carragher, B.; Veesler, D.
Subnanometre-resolution electron cryomicroscopy structure of a heterodimeric ABC exporter
Nature
2014
Kim, J.; Wu, S.; Tomasiak, T. M.; Mergel, C.; Winter, M. B.; Stiller, S. B.; Robles-Colmanares, Y.; Stroud, R. M.; Tampé, R.; Craik, C. S.; Cheng, Y.
Visualization of ATP synthase dimers in mitochondria by electron cryo-tomography
Journal of Visual Experiments: JOVE
2014
Davies, K. M.; Daum, B.; Gold, V. A.; Mühleip, A. W.; Brandt, T.; Blum, T. B.; Mills, D. J.; Kühlbrandt, W.
Molecular imprinting as a signal-activation mechanism of the viral RNA sensor RIG-I
Molecular Cell
2014
Wu, B.; Peisley, A.; Tetrault, D.; Li, Z.; Egelman, E. H.; Magor, K. E.; Walz, T.; Penczek, P. A.;Hur, S.
Structure of β-galactosidase at 3.2-Å resolution obtained by cryo-electron microscopy
Proceedings of the National Acadamy of Sciences
2014
Bartesaghi, A.; Matthies, D.; Banerjee, S.; Merk, A.; Subramaniam, S.
Single particle electron cryo-microscopy of a mammalian ion channel
Current Opinion in Structural Biology
2014
Liao, M.; Cao, E.; Julius, D.; Cheng, Y.
M-free: Scoring the reference bias in sub-tomogram averaging and template matching
Journal of Structural Biology
2014
Yu, Z.; Frangakis, A. S.
Visualizing active membrane protein complexes by electron cryotomography
Nature Communications
2014
Gold, V. A. M.; Ieva, R.; Walter, A.; Pfanner, N.; van der Laan, M.; Kühlbrandt, W.
CTER-rapid estimation of CTF parameters with error assessment
Ultramicroscopy
2014
Penczek, P. A.; Fang, J.; Li, X.; Cheng, Y.; Loerke, J.; Spahn, C. M. T.
Journal of Analytical Science and Technology
2014
Cho, H. J.; Hyun, J. K.; Kim, J. G.; Jeong, H. S.; Park, H. N.; You, D. J.; Jung, H. S.
Architecture and host interface of environmental chlamydiae revealed by electron cryotomography
Environmental Microbiology
2014
Pilhofer, M.; Aistleitner, K.; Ladinsky, M. S.; König, L.; Horn, M.; Jensen, G. J.
Structural basis for the prion-like MAVS filaments in antiviral innate immunity
eLife
2014
Xu, H.; He, X.; Zheng, H.; Huang, L. J.; Hou, F.; Yu, Z.; de la Cruz, M. J.; Borkowski, B.; Zhang, X.; Chen, Z. J.; Jiang, Q. X.
Cryo-EM structure of a fully glycosylated soluble cleaved HIV-1 envelope trimer
Science
2013
Lyumkis, D.; Julien, J. P.; de Val, N.; Cupo, A.; Potter, C. S.; Klasse, P. J.; Burton, D. R.; Sanders, R. W.; Moore, J. P.; Carragher, B.; Wilson, I. A.; Ward, A. B.
TRPV1 structures in distinct confirmations reveal activation mechanisms
Nature
2013
Cao, E.; Liao, M.; Cheng, Y.; Julius, D.
Maximizing the potential of electron cryomicroscopy data collected using direct detectors
Journal of Structural Biology
2013
Veesler, D.; Campbell, M. G.; Cheng, A.; Fu, C.; Murez, Z.; Johnson, J. E.; Potter, C. S.; Carragher, B.
Polyphosphate storage during sporulation in the gram-negative bacterium acetonema iongum
Journal of Bacteriology
2013
Tocheva, E. I.; Dekas, A. E.; McGlynn, S. E.; Morris, D.; Orphan, V. J.; Jensen, G. J.
Peptidoglycan transformations during Bacillus subtilis sporulation
Molecular Microbiology
2013
Tocheva, E. I.; López-Garrido, J.; Hughes, H. V.; Fredlund ,J.; Kuru, E.; Vannieuwenhze, M. S.; Brun, Y. V.; Pogliano, K.; Jensen. G. J.
Compression deformation of single-crystal Pt3Al with the L12 structure
MRS Proceedings
2013
Hasegawa, Y.; Okamoto, N. L.; Inui, H.
Single particle cryo-electron microscopy and 3-D reconstruction of viruses
Electron Microscopy
2012
Guo, F.; Jiang, W.
应用
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Breaking the molecular weight barriers for cryo-electron microscopy |
|
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| 冷冻电子显微术要求为显微镜采用特殊环境 |
相关材料
Nyquist 频率
剂量分割和运动校正
使用计数和超分辨率提高 DQE
| Modulation transfer function (MTF) curves | ||
|---|---|---|
| 200 kV | 300 kV | |
|
K3 |
CDS | CDS |
| Standard | Standard | |
| K2 | Standard | Standard |