IDEAS home Printed from https://ideas.repec.org/a/nat/natcom/v15y2024i1d10.1038_s41467-024-50462-2.html
   My bibliography  Save this article

Versatile nanobody-based approach to image, track and reconstitute functional Neurexin-1 in vivo

Author

Listed:
  • Rosario Vicidomini

    (NIH)

  • Saumitra Dey Choudhury

    (NIH
    New Delhi)

  • Tae Hee Han

    (NIH)

  • Tho Huu Nguyen

    (NIH)

  • Peter Nguyen

    (NIH)

  • Felipe Opazo

    (University Medical Center Göttingen
    NanoTag Biotechnologies GmbH)

  • Mihaela Serpe

    (NIH)

Abstract

Neurexins are key adhesion proteins that coordinate extracellular and intracellular synaptic components. Nonetheless, the low abundance of these multidomain proteins has complicated any localization and structure-function studies. Here we combine an ALFA tag (AT)/nanobody (NbALFA) tool with classic genetics, cell biology and electrophysiology to examine the distribution and function of the Drosophila Nrx-1 in vivo. We generate full-length and ΔPDZ ALFA-tagged Nrx-1 variants and find that the PDZ binding motif is key to Nrx-1 surface expression. A PDZ binding motif provided in trans, via genetically encoded cytosolic NbALFA-PDZ chimera, fully restores the synaptic localization and function of NrxΔPDZ-AT. Using cytosolic NbALFA-mScarlet intrabody, we achieve compartment-specific detection of endogenous Nrx-1, track live Nrx-1 transport along the motor neuron axons, and demonstrate that Nrx-1 co-migrates with Rab2-positive vesicles. Our findings illustrate the versatility of the ALFA system and pave the way towards dissecting functional domains of complex proteins in vivo.

Suggested Citation

  • Rosario Vicidomini & Saumitra Dey Choudhury & Tae Hee Han & Tho Huu Nguyen & Peter Nguyen & Felipe Opazo & Mihaela Serpe, 2024. "Versatile nanobody-based approach to image, track and reconstitute functional Neurexin-1 in vivo," Nature Communications, Nature, vol. 15(1), pages 1-17, December.
  • Handle: RePEc:nat:natcom:v:15:y:2024:i:1:d:10.1038_s41467-024-50462-2
    DOI: 10.1038/s41467-024-50462-2
    as

    Download full text from publisher

    File URL: https://www.nature.com/articles/s41467-024-50462-2
    File Function: Abstract
    Download Restriction: no

    File URL: https://libkey.io/10.1038/s41467-024-50462-2?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. Thomas C. Südhof, 2008. "Neuroligins and neurexins link synaptic function to cognitive disease," Nature, Nature, vol. 455(7215), pages 903-911, October.
    2. Olena Riabinina & Darya Task & Elizabeth Marr & Chun-Chieh Lin & Robert Alford & David A. O'Brochta & Christopher J. Potter, 2016. "Organization of olfactory centres in the malaria mosquito Anopheles gambiae," Nature Communications, Nature, vol. 7(1), pages 1-12, December.
    3. Karzan Muhammad & Suneel Reddy-Alla & Jan H Driller & Dietmar Schreiner & Ulises Rey & Mathias A. Böhme & Christina Hollmann & Niraja Ramesh & Harald Depner & Janine Lützkendorf & Tanja Matkovic & Tor, 2015. "Presynaptic spinophilin tunes neurexin signalling to control active zone architecture and function," Nature Communications, Nature, vol. 6(1), pages 1-15, December.
    4. Hansjörg Götzke & Markus Kilisch & Markel Martínez-Carranza & Shama Sograte-Idrissi & Abirami Rajavel & Thomas Schlichthaerle & Niklas Engels & Ralf Jungmann & Pål Stenmark & Felipe Opazo & Steffen Fr, 2019. "The ALFA-tag is a highly versatile tool for nanobody-based bioscience applications," Nature Communications, Nature, vol. 10(1), pages 1-12, December.
    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. Tales V. Pascini & Yeong Je Jeong & Wei Huang & Zarna R. Pala & Juliana M. Sá & Michael B. Wells & Christopher Kizito & Brendan Sweeney & Thiago L. Alves e Silva & Deborah J. Andrew & Marcelo Jacobs-­, 2022. "Transgenic Anopheles mosquitoes expressing human PAI-1 impair malaria transmission," Nature Communications, Nature, vol. 13(1), pages 1-16, December.
    2. Leo Kiss & Tyler Rhinesmith & Jakub Luptak & Claire F. Dickson & Jonas Weidenhausen & Shannon Smyly & Ji-Chun Yang & Sarah L. Maslen & Irmgard Sinning & David Neuhaus & Dean Clift & Leo C. James, 2023. "Trim-Away ubiquitinates and degrades lysine-less and N-terminally acetylated substrates," Nature Communications, Nature, vol. 14(1), pages 1-17, December.
    3. Xudong Chen & Min Xie & Sensen Zhang & Marta Monguió-Tortajada & Jian Yin & Chang Liu & Youqi Zhang & Maeva Delacrétaz & Mingyue Song & Yixue Wang & Lin Dong & Qiang Ding & Boda Zhou & Xiaolin Tian & , 2023. "Structural basis for recruitment of TASL by SLC15A4 in human endolysosomal TLR signaling," Nature Communications, Nature, vol. 14(1), pages 1-13, December.
    4. Xiansha Xiao & Allison Fay & Pablo Santos Molina & Amanda Kovach & Michael S. Glickman & Huilin Li, 2024. "Structure of the M. tuberculosis DnaK−GrpE complex reveals how key DnaK roles are controlled," Nature Communications, Nature, vol. 15(1), pages 1-16, December.
    5. Kevin Wu & Samuel Itskanov & Diane L. Lynch & Yuanyuan Chen & Aasha Turner & James C. Gumbart & Eunyong Park, 2024. "Substrate recognition mechanism of the endoplasmic reticulum-associated ubiquitin ligase Doa10," Nature Communications, Nature, vol. 15(1), pages 1-18, December.
    6. Pranjul Singh & Shefali Goyal & Smith Gupta & Sanket Garg & Abhinav Tiwari & Varad Rajput & Alexander Shakeel Bates & Arjit Kant Gupta & Nitin Gupta, 2023. "Combinatorial encoding of odors in the mosquito antennal lobe," Nature Communications, Nature, vol. 14(1), pages 1-19, December.
    7. Irina Shlosman & Elayne M. Fivenson & Morgan S. A. Gilman & Tyler A. Sisley & Suzanne Walker & Thomas G. Bernhardt & Andrew C. Kruse & Joseph J. Loparo, 2023. "Allosteric activation of cell wall synthesis during bacterial growth," 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:15:y:2024:i:1:d:10.1038_s41467-024-50462-2. 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.