IDEAS home Printed from https://ideas.repec.org/a/nat/natcom/v16y2025i1d10.1038_s41467-025-59674-6.html
   My bibliography  Save this article

Single-molecule visualization of ATP-induced dynamics of the subunit composition of an ECF transporter complex under turnover conditions

Author

Listed:
  • Solène N. Lefebvre

    (University of Groningen)

  • Mark Nijland

    (University of Groningen)

  • Ivan Maslov

    (University of Groningen)

  • Dirk J. Slotboom

    (University of Groningen)

Abstract

Energy-Coupling Factor (ECF) transporters are ATP-binding cassette (ABC) transporters essential for uptake of vitamins and cofactors in prokaryotes. They have been linked to pathogen virulence and are potential targets for antimicrobials. ECF transporters have been proposed to use a unique transport mechanism where a substrate-translocating subunit (S-component) dynamically associates with and dissociates from an ATP-hydrolyzing motor (ECF module). This model is contentious, because it is based largely on experimental conditions without compartments or continuous bilayers. Here, we used single-molecule spectroscopy to investigate the conformational dynamics of the vitamin B12 transporter ECF-CbrT in membranes under vectorial transport conditions. We observed ATP hydrolysis-dependent dissociation of the S-component CbrT from, and re-association with the ECF module, in absence and presence of vitamin B12 consistent with futile ATP hydrolysis activity. The single-molecule spectroscopy experiments suggest that S-component expulsion from and re-association with the ECF module are an integral part of the translocation mechanism.

Suggested Citation

  • Solène N. Lefebvre & Mark Nijland & Ivan Maslov & Dirk J. Slotboom, 2025. "Single-molecule visualization of ATP-induced dynamics of the subunit composition of an ECF transporter complex under turnover conditions," Nature Communications, Nature, vol. 16(1), pages 1-12, December.
    • Handle: RePEc:nat:natcom:v:16:y:2025:i:1:d:10.1038_s41467-025-59674-6
    DOI: 10.1038/s41467-025-59674-6
    as

    Download full text from publisher

    File URL: https://www.nature.com/articles/s41467-025-59674-6
    File Function: Abstract
    Download Restriction: no
    File URL: https://libkey.io/10.1038/s41467-025-59674-6?utm_source=ideas
    LibKey link: if access is restricted and if your library uses this service, LibKey will redirect you to where you can use your library subscription to access this item
    ---><---

    References listed on IDEAS

    as
    1. Antonino Ingargiola & Eitan Lerner & SangYoon Chung & Shimon Weiss & Xavier Michalet, 2016. "FRETBursts: An Open Source Toolkit for Analysis of Freely-Diffusing Single-Molecule FRET," PLOS ONE, Public Library of Science, vol. 11(8), pages 1-27, August.
    2. S. Rempel & E. Colucci & J. W. Gier & A. Guskov & D. J. Slotboom, 2018. "Cysteine-mediated decyanation of vitamin B12 by the predicted membrane transporter BtuM," Nature Communications, Nature, vol. 9(1), pages 1-8, December.
    3. Ignacio Faustino & Haleh Abdizadeh & Paulo C. T. Souza & Aike Jeucken & Weronika K. Stanek & Albert Guskov & Dirk J. Slotboom & Siewert J. Marrink, 2020. "Membrane mediated toppling mechanism of the folate energy coupling factor transporter," Nature Communications, Nature, vol. 11(1), pages 1-9, December.
    4. Chancievan Thangaratnarajah & Mark Nijland & Luís Borges-Araújo & Aike Jeucken & Jan Rheinberger & Siewert J. Marrink & Paulo C. T. Souza & Cristina Paulino & Dirk J. Slotboom, 2023. "Expulsion mechanism of the substrate-translocating subunit in ECF transporters," Nature Communications, Nature, vol. 14(1), pages 1-14, December.
    5. Joris M. H. Goudsmits & Dirk Jan Slotboom & Antoine M. Oijen, 2017. "Single-molecule visualization of conformational changes and substrate transport in the vitamin B12 ABC importer BtuCD-F," Nature Communications, Nature, vol. 8(1), pages 1-10, December.
    6. Lotteke J. Y. M. Swier & Albert Guskov & Dirk J. Slotboom, 2016. "Structural insight in the toppling mechanism of an energy-coupling factor transporter," Nature Communications, Nature, vol. 7(1), pages 1-11, April.
    7. Youichi Naoe & Nozomi Nakamura & Akihiro Doi & Mia Sawabe & Hiro Nakamura & Yoshitsugu Shiro & Hiroshi Sugimoto, 2016. "Crystal structure of bacterial haem importer complex in the inward-facing conformation," Nature Communications, Nature, vol. 7(1), pages 1-11, December.
    8. Mark Nijland & Solène N. Lefebvre & Chancievan Thangaratnarajah & Dirk J. Slotboom, 2024. "Bidirectional ATP-driven transport of cobalamin by the mycobacterial ABC transporter BacA," Nature Communications, Nature, vol. 15(1), pages 1-9, December.
    9. Gabriel A. Fitzgerald & Daniel S. Terry & Audrey L. Warren & Matthias Quick & Jonathan A. Javitch & Scott C. Blanchard, 2019. "Quantifying secondary transport at single-molecule resolution," Nature, Nature, vol. 575(7783), pages 528-534, November.
    10. S. Rempel & C. Gati & M. Nijland & C. Thangaratnarajah & A. Karyolaimos & J. W. Gier & A. Guskov & D. J. Slotboom, 2020. "A mycobacterial ABC transporter mediates the uptake of hydrophilic compounds," Nature, Nature, vol. 580(7803), pages 409-412, April.
    Full references (including those not matched with items on IDEAS)

    Most related items

    These are the items that most often cite the same works as this one and are cited by the same works as this one.
    1. Mark Nijland & Solène N. Lefebvre & Chancievan Thangaratnarajah & Dirk J. Slotboom, 2024. "Bidirectional ATP-driven transport of cobalamin by the mycobacterial ABC transporter BacA," Nature Communications, Nature, vol. 15(1), pages 1-9, December.
    2. Chancievan Thangaratnarajah & Mark Nijland & Luís Borges-Araújo & Aike Jeucken & Jan Rheinberger & Siewert J. Marrink & Paulo C. T. Souza & Cristina Paulino & Dirk J. Slotboom, 2023. "Expulsion mechanism of the substrate-translocating subunit in ECF transporters," Nature Communications, Nature, vol. 14(1), pages 1-14, December.
    3. Wanfeng Xu & Yun Wang & Shuang Cui & Qiuling Zheng & Yanghao Lin & Qingqing Cui & Yuxin Xie & Yuming Zeng & Chuan Zhang & Yujie Li & Xin Jin & Minna Qin & Huiyong Sun & Haiping Hao & Lijuan Cao, 2025. "Methylcobalamin protects against liver failure via engaging gasdermin E," Nature Communications, Nature, vol. 16(1), pages 1-16, December.
    4. Jere P. Segrest & Chongren Tang & Hyun D. Song & Martin K. Jones & W. Sean Davidson & Stephen G. Aller & Jay W. Heinecke, 2022. "ABCA1 is an extracellular phospholipid translocase," Nature Communications, Nature, vol. 13(1), pages 1-12, December.
    5. Lin Yu & Xin Xu & Wan-Zhen Chua & Hao Feng & Zheng Ser & Kai Shao & Jian Shi & Yumei Wang & Zongli Li & Radoslaw M. Sobota & Lok-To Sham & Min Luo, 2023. "Structural basis of peptide secretion for Quorum sensing by ComA," Nature Communications, Nature, vol. 14(1), pages 1-17, December.
    6. Christian Gebhardt & Pascal Bawidamann & Anna-Katharina Spring & Robin Schenk & Konstantin Schütze & Gabriel G. Moya Muñoz & Nicolas D. Wendler & Douglas A. Griffith & Jan Lipfert & Thorben Cordes, 2025. "Labelizer: systematic selection of protein residues for covalent fluorophore labeling," Nature Communications, Nature, vol. 16(1), pages 1-16, December.
    7. Katherine L. Fenn & Jim E. Horne & Joel A. Crossley & Nils Böhringer & Romany J. Horne & Till F. Schäberle & Antonio N. Calabrese & Sheena E. Radford & Neil A. Ranson, 2024. "Outer membrane protein assembly mediated by BAM-SurA complexes," Nature Communications, Nature, vol. 15(1), pages 1-17, December.
    8. Markus Götz & Anders Barth & Søren S. -R. Bohr & Richard Börner & Jixin Chen & Thorben Cordes & Dorothy A. Erie & Christian Gebhardt & Mélodie C. A. S. Hadzic & George L. Hamilton & Nikos S. Hatzakis , 2024. "Reply to: On the statistical foundation of a recent single molecule FRET benchmark," Nature Communications, Nature, vol. 15(1), pages 1-4, December.
    9. Antonino Ingargiola & Eitan Lerner & SangYoon Chung & Francesco Panzeri & Angelo Gulinatti & Ivan Rech & Massimo Ghioni & Shimon Weiss & Xavier Michalet, 2017. "Multispot single-molecule FRET: High-throughput analysis of freely diffusing molecules," PLOS ONE, Public Library of Science, vol. 12(4), pages 1-27, April.
    10. Paul David Harris & Alessandra Narducci & Christian Gebhardt & Thorben Cordes & Shimon Weiss & Eitan Lerner, 2022. "Multi-parameter photon-by-photon hidden Markov modeling," Nature Communications, Nature, vol. 13(1), pages 1-12, December.
    11. Hyunju Cho & Yumeng Liu & SangYoon Chung & Sowmya Chandrasekar & Shimon Weiss & Shu-ou Shan, 2024. "Dynamic stability of Sgt2 enables selective and privileged client handover in a chaperone triad," Nature Communications, Nature, vol. 15(1), pages 1-16, December.
    12. Michael C. Owens & Hui Shen & Amber Yanas & Maria Saraí Mendoza-Figueroa & Ellen Lavorando & Xiaoyu Wei & Him Shweta & Hsin-Yao Tang & Yale E. Goldman & Kathy Fange Liu, 2024. "Specific catalytically impaired DDX3X mutants form sexually dimorphic hollow condensates," Nature Communications, Nature, vol. 15(1), pages 1-17, December.
    13. Pujun Xie & Yan Li & Gaëlle Lamon & Huihui Kuang & Da-Neng Wang & Nathaniel J. Traaseth, 2025. "A fiducial-assisted strategy compatible with resolving small MFS transporter structures in multiple conformations using cryo-EM," Nature Communications, Nature, vol. 16(1), pages 1-17, December.
    14. Bob Schiffrin & Joel A. Crossley & Martin Walko & Jonathan M. Machin & G. Nasir Khan & Iain W. Manfield & Andrew J. Wilson & David J. Brockwell & Tomas Fessl & Antonio N. Calabrese & Sheena E. Radford, 2024. "Dual client binding sites in the ATP-independent chaperone SurA," Nature Communications, Nature, vol. 15(1), pages 1-16, December.

    More about this item

    Statistics

    Access and download statistics

    Corrections

    All material on this site has been provided by the respective publishers and authors. You can help correct errors and omissions. When requesting a correction, please mention this item's handle: RePEc:nat:natcom:v:16:y:2025:i:1:d:10.1038_s41467-025-59674-6. See general information about how to correct material in RePEc.

    If you have authored this item and are not yet registered with RePEc, we encourage you to do it here. This allows to link your profile to this item. It also allows you to accept potential citations to this item that we are uncertain about.

    If CitEc recognized a bibliographic reference but did not link an item in RePEc to it, you can help with this form .

    If you know of missing items citing this one, you can help us creating those links by adding the relevant references in the same way as above, for each refering item. If you are a registered author of this item, you may also want to check the "citations" tab in your RePEc Author Service profile, as there may be some citations waiting for confirmation.

    For technical questions regarding this item, or to correct its authors, title, abstract, bibliographic or download information, contact: Sonal Shukla or Springer Nature Abstracting and Indexing (email available below). General contact details of provider: http://www.nature.com .

    Please note that corrections may take a couple of weeks to filter through the various RePEc services.

    IDEAS is a RePEc service. RePEc uses bibliographic data supplied by the respective publishers.