IDEAS home Printed from https://ideas.repec.org/a/nat/natcom/v16y2025i1d10.1038_s41467-025-56629-9.html
   My bibliography  Save this article

Mutant glycosidases for labeling sialoglycans with high specificity and affinity

Author

Listed:
  • Shuyu Liang

    (Peking University
    Peking University)

  • Qi Tang

    (Peking University
    Peking University)

  • Xunzi Guo

    (Peking University
    Peking University)

  • Zi’an Li

    (Peking University
    Peking University)

  • Yilan Guo

    (Peking University
    Peking University)

  • Jinghan Chang

    (Peking University
    Peking University)

  • Bo Cheng

    (Peking University
    Peking University)

  • Qitao Song

    (Peking University
    Peking University)

  • Jiayu Sun

    (Peking University
    Peking University)

  • Peng Dai

    (Peking University
    Peking University
    Peking University
    Peking University)

  • Xing Chen

    (Peking University
    Peking University
    Peking University
    Peking University)

Abstract

Affinity labeling of biomacromolecules is vital for bioimaging and functional studies. However, affinity probes recognizing glycans with high specificity remain scarce. Here we report the development of glycan recombinant affinity binders (GRABs) based on mutant bacterial sialidases, which are enzymatically inactive but preserve stringent specificity for sialoglycan substrates. By mutating a key catalytic residue of Streptococcus pneumoniae neuraminidase A (SpNanA) and Ruminococcus gnavus neuraminidase H (RgNanH), we develop GRAB-Sia and GRAB-Sia3 recognizing total sialoglycans and α2,3-sialosides, respectively. The GRABs exhibit strict substrate and linkage specificity, and tetramerization with streptavidin substantially increases their avidity. The GRABs and tetrameric GRABs (tetra-GRABs) are effective tools for probing sialoglycans in immunoblotting, flow cytometry, immunoprecipitation, and fluorescence imaging. Furthermore, multiplex analysis with tetra-GRABs uncovers spatially distinct sialoglycans in the various mouse organs. This work provides a versatile toolkit for labeling and analyzing sialoglycans with high specificity, sensitivity, and convenience.

Suggested Citation

  • Shuyu Liang & Qi Tang & Xunzi Guo & Zi’an Li & Yilan Guo & Jinghan Chang & Bo Cheng & Qitao Song & Jiayu Sun & Peng Dai & Xing Chen, 2025. "Mutant glycosidases for labeling sialoglycans with high specificity and affinity," Nature Communications, Nature, vol. 16(1), pages 1-14, December.
  • Handle: RePEc:nat:natcom:v:16:y:2025:i:1:d:10.1038_s41467-025-56629-9
    DOI: 10.1038/s41467-025-56629-9
    as

    Download full text from publisher

    File URL: https://www.nature.com/articles/s41467-025-56629-9
    File Function: Abstract
    Download Restriction: no

    File URL: https://libkey.io/10.1038/s41467-025-56629-9?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. Emily Rodrigues & Jaesoo Jung & Heajin Park & Caleb Loo & Sepideh Soukhtehzari & Elena N. Kitova & Fahima Mozaneh & Gour Daskhan & Edward N. Schmidt & Vivian Aghanya & Susmita Sarkar & Laura Streith &, 2020. "A versatile soluble siglec scaffold for sensitive and quantitative detection of glycan ligands," Nature Communications, Nature, vol. 11(1), pages 1-13, December.
    2. Louise E. Tailford & C. David Owen & John Walshaw & Emmanuelle H. Crost & Jemma Hardy-Goddard & Gwenaelle Le Gall & Willem M. de Vos & Garry L. Taylor & Nathalie Juge, 2015. "Discovery of intramolecular trans-sialidases in human gut microbiota suggests novel mechanisms of mucosal adaptation," Nature Communications, Nature, vol. 6(1), pages 1-12, November.
    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. Chih-Lan Lin & Mirat Sojitra & Eric J. Carpenter & Ellen S. Hayhoe & Susmita Sarkar & Elizabeth A. Volker & Chao Wang & Duong T. Bui & Loretta Yang & John S. Klassen & Peng Wu & Matthew S. Macauley & , 2023. "Chemoenzymatic synthesis of genetically-encoded multivalent liquid N-glycan arrays," Nature Communications, Nature, vol. 14(1), pages 1-15, December.
    2. Janneke Elzinga & Yoshiki Narimatsu & Noortje Haan & Henrik Clausen & Willem M. Vos & Hanne L. P. Tytgat, 2024. "Binding of Akkermansia muciniphila to mucin is O-glycan specific," Nature Communications, Nature, vol. 15(1), pages 1-10, December.
    3. Edward N. Schmidt & Dimitra Lamprinaki & Kelli A. McCord & Maju Joe & Mirat Sojitra & Ayk Waldow & Jasmine Nguyen & John Monyror & Elena N. Kitova & Fahima Mozaneh & Xue Yan Guo & Jaesoo Jung & Jhon R, 2023. "Siglec-6 mediates the uptake of extracellular vesicles through a noncanonical glycolipid binding pocket," Nature Communications, Nature, vol. 14(1), pages 1-17, December.
    4. Maria Pia Lenza & Leire Egia-Mendikute & Asier Antoñana-Vildosola & Cátia O. Soares & Helena Coelho & Francisco Corzana & Alexandre Bosch & Prodhi Manisha & Jon Imanol Quintana & Iker Oyenarte & Luca , 2023. "Structural insights into Siglec-15 reveal glycosylation dependency for its interaction with T cells through integrin CD11b," Nature Communications, Nature, vol. 14(1), pages 1-14, 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_s41467-025-56629-9. 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.