ACS Publications. Most Trusted. Most Cited. Most Read
Modular Imaging Scaffold for Single-Particle Electron Microscopy
My Activity
    Article

    Modular Imaging Scaffold for Single-Particle Electron Microscopy
    Click to copy article linkArticle link copied!

    • Nesrine Aissaoui
      Nesrine Aissaoui
      Centre de Biochimie Structurale, CNRS UMR 5048, INSERM U1054, F-34000 Montpellier, France
      Université de Montpellier, F-34000 Montpellier, France
    • Josephine Lai-Kee-Him
      Josephine Lai-Kee-Him
      Centre de Biochimie Structurale, CNRS UMR 5048, INSERM U1054, F-34000 Montpellier, France
      Université de Montpellier, F-34000 Montpellier, France
    • Allan Mills
      Allan Mills
      Centre de Biochimie Structurale, CNRS UMR 5048, INSERM U1054, F-34000 Montpellier, France
      Université de Montpellier, F-34000 Montpellier, France
      More by Allan Mills
    • Nathalie Declerck
      Nathalie Declerck
      Centre de Biochimie Structurale, CNRS UMR 5048, INSERM U1054, F-34000 Montpellier, France
      Université de Montpellier, F-34000 Montpellier, France
      Departement MICA, INRA, 78352 Jouy-en-Josas, France
    • Zakia Morichaud
      Zakia Morichaud
      Université de Montpellier, F-34000 Montpellier, France
      IRIM, CNRS, Université Montpellier, 1919 Route de Mende, 34293 Montpellier, France
    • Konstantin Brodolin
      Konstantin Brodolin
      Université de Montpellier, F-34000 Montpellier, France
      IRIM, CNRS, Université Montpellier, 1919 Route de Mende, 34293 Montpellier, France
    • Sonia Baconnais
      Sonia Baconnais
      Signalisations, Noyaux et Innovations en Cancérologie, UMR 8126, CNRS, Université Paris-Sud, Gustave Roussy, Université Paris-Saclay, 94800 Villejuif, France
    • Eric Le Cam
      Eric Le Cam
      Signalisations, Noyaux et Innovations en Cancérologie, UMR 8126, CNRS, Université Paris-Sud, Gustave Roussy, Université Paris-Saclay, 94800 Villejuif, France
      More by Eric Le Cam
    • Jean Baptiste Charbonnier
      Jean Baptiste Charbonnier
      Université Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), 91198 Gif-sur-Yvette, France
    • Rémy Sounier
      Rémy Sounier
      Université de Montpellier, F-34000 Montpellier, France
      Institut de Génomique Fonctionnelle, CNRS UMR 5203, INSERM U1191, F-34000 Montpellier, France
    • Sébastien Granier
      Sébastien Granier
      Université de Montpellier, F-34000 Montpellier, France
      Institut de Génomique Fonctionnelle, CNRS UMR 5203, INSERM U1191, F-34000 Montpellier, France
    • Virginie Ropars*
      Virginie Ropars
      Université Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), 91198 Gif-sur-Yvette, France
      *E-mail: [email protected]
    • Patrick Bron*
      Patrick Bron
      Centre de Biochimie Structurale, CNRS UMR 5048, INSERM U1054, F-34000 Montpellier, France
      Université de Montpellier, F-34000 Montpellier, France
      *E-mail: [email protected]
      More by Patrick Bron
    • Gaetan Bellot*
      Gaetan Bellot
      Centre de Biochimie Structurale, CNRS UMR 5048, INSERM U1054, F-34000 Montpellier, France
      Université de Montpellier, F-34000 Montpellier, France
      *E-mail: [email protected]
    Other Access OptionsSupporting Information (1)

    ACS Nano

    Cite this: ACS Nano 2021, 15, 3, 4186–4196
    Click to copy citationCitation copied!
    https://doi.org/10.1021/acsnano.0c05113
    Published February 15, 2021
    Copyright © 2021 American Chemical Society

    Abstract

    Click to copy section linkSection link copied!
    Abstract Image

    Technological breakthroughs in electron microscopy (EM) have made it possible to solve structures of biological macromolecular complexes and to raise novel challenges, specifically related to sample preparation and heterogeneous macromolecular assemblies such as DNA–protein, protein–protein, and membrane protein assemblies. Here, we built a V-shaped DNA origami as a scaffolding molecular system to template proteins at user-defined positions in space. This template positions macromolecular assemblies of various sizes, juxtaposes combinations of biomolecules into complex arrangements, isolates biomolecules in their active state, and stabilizes membrane proteins in solution. In addition, the design can be engineered to tune DNA mechanical properties by exerting a controlled piconewton (pN) force on the molecular system and thus adapted to characterize mechanosensitive proteins. The binding site can also be specifically customized to accommodate the protein of interest, either interacting spontaneously with DNA or through directed chemical conjugation, increasing the range of potential targets for single-particle EM investigation. We assessed the applicability for five different proteins. Finally, as a proof of principle, we used RNAP protein to validate the approach and to explore the compatibility of the template with cryo-EM sample preparation.

    Copyright © 2021 American Chemical Society

    Read this article

    To access this article, please review the available access options below.

    Get instant access

    Purchase Access

    Read this article for 48 hours. Check out below using your ACS ID or as a guest.

    Recommended

    Access through Your Institution

    You may have access to this article through your institution.

    Your institution does not have access to this content. Add or change your institution or let them know you’d like them to include access.

    Supporting Information

    Click to copy section linkSection link copied!

    The Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/acsnano.0c05113.

    • Design and characterization of DNA origami structures as well as additional TEM images of the different protein attachments to DNA origami binding sites; the model applied to estimate the force generated on the RNAP-DNA binding domain; list of all oligonucleotides (PDF)

    Terms & Conditions

    Most electronic Supporting Information files are available without a subscription to ACS Web Editions. Such files may be downloaded by article for research use (if there is a public use license linked to the relevant article, that license may permit other uses). Permission may be obtained from ACS for other uses through requests via the RightsLink permission system: http://pubs.acs.org/page/copyright/permissions.html.

    Cited By

    Click to copy section linkSection link copied!
    Citation Statements
    Explore this article's citation statements on scite.ai

    This article is cited by 9 publications.

    1. Yuanfang Chen, Yiqian Huang, Yuhe R. Yang. DNA Nanotags for Multiplexed Single-Particle Electron Microscopy and In Situ Electron Cryotomography. JACS Au 2025, 5 (1) , 17-27. https://doi.org/10.1021/jacsau.4c00986
    2. Nesrine Aissaoui, Allan Mills, Josephine Lai-Kee-Him, Nicolas Triomphe, Quentin Cece, Christine Doucet, Anne Bonhoure, Michel Vidal, Yonggang Ke, Gaetan Bellot. Free-Standing DNA Origami Superlattice to Facilitate Cryo-EM Visualization of Membrane Vesicles. Journal of the American Chemical Society 2024, 146 (19) , 12925-12932. https://doi.org/10.1021/jacs.3c07328
    3. Kun Zhou, Minhwan Chung, Jing Cheng, John T. Powell, Qi Yan, Jun Liu, Yong Xiong, Martin A. Schwartz, Chenxiang Lin. DNA nanodevice for analysis of force-activated protein extension and interactions. 2024https://doi.org/10.1101/2024.10.25.620262
    4. Ashwin Karthick Natarajan, Joonas Ryssy, Anton Kuzyk. A DNA origami-based device for investigating DNA bending proteins by transmission electron microscopy. Nanoscale 2023, 15 (7) , 3212-3218. https://doi.org/10.1039/D2NR05366G
    5. Chun Xie, Yingxin Hu, Zhekun Chen, Kuiting Chen, Linqiang Pan. Tuning curved DNA origami structures through mechanical design and chemical adducts. Nanotechnology 2022, 33 (40) , 405603. https://doi.org/10.1088/1361-6528/ac7d62
    6. Brandon Huntington, Lingyun Zhao, Patrick Bron, Umar F. Shahul Hameed, Stefan T. Arold, Bilal M. Qureshi. Thicker Ice Improves the Integrity and Angular Distribution of CDC48A Hexamers on Cryo-EM Grids. Frontiers in Molecular Biosciences 2022, 9 https://doi.org/10.3389/fmolb.2022.890390
    7. Koen Wentinck, Christos Gogou, Dimphna H. Meijer. Putting on molecular weight: Enabling cryo-EM structure determination of sub-100-kDa proteins. Current Research in Structural Biology 2022, 4 , 332-337. https://doi.org/10.1016/j.crstbi.2022.09.005
    8. Muchen Pan, Jiye Shi, Lihua Wang, Chunhai Fan, Xiaoguo Liu. Cryogenic Electron Microscopy for Resolving DNA Nanostructures and Their Complexes. Small Structures 2021, 2 (10) https://doi.org/10.1002/sstr.202100053
    9. Qian Tang, Da Han. Obtaining Precise Molecular Information via DNA Nanotechnology. Membranes 2021, 11 (9) , 683. https://doi.org/10.3390/membranes11090683
    10. Sayma Zahid, Murielle Seif El Dahan, Florence Iehl, Paloma Fernandez-Varela, Marie-Helene Le Du, Virginie Ropars, Jean Baptiste Charbonnier. The Multifaceted Roles of Ku70/80. International Journal of Molecular Sciences 2021, 22 (8) , 4134. https://doi.org/10.3390/ijms22084134

    ACS Nano

    Cite this: ACS Nano 2021, 15, 3, 4186–4196
    Click to copy citationCitation copied!
    https://doi.org/10.1021/acsnano.0c05113
    Published February 15, 2021
    Copyright © 2021 American Chemical Society

    Article Views

    1616

    Altmetric

    -

    Citations

    Learn about these metrics

    Article Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.

    Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.

    The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated.