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Bioorthogonal labeling of transmembrane proteins with non-canonical amino acids unveils masked epitopes in live neurons

Author

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  • Diogo Bessa-Neto

    (University of Bordeaux, CNRS, Interdisciplinary Institute for Neuroscience, IINS, UMR 5297)

  • Gerti Beliu

    (University of Würzburg, Biocenter, Am Hubland
    Rudolf Virchow Center for Integrative and Translational Bioimaging, University of Wuerzburg)

  • Alexander Kuhlemann

    (University of Würzburg, Biocenter, Am Hubland)

  • Valeria Pecoraro

    (University of Bordeaux, CNRS, Interdisciplinary Institute for Neuroscience, IINS, UMR 5297)

  • Sören Doose

    (University of Würzburg, Biocenter, Am Hubland)

  • Natacha Retailleau

    (University of Bordeaux, CNRS, Interdisciplinary Institute for Neuroscience, IINS, UMR 5297)

  • Nicolas Chevrier

    (University of Bordeaux, CNRS, Interdisciplinary Institute for Neuroscience, IINS, UMR 5297)

  • David Perrais

    (University of Bordeaux, CNRS, Interdisciplinary Institute for Neuroscience, IINS, UMR 5297)

  • Markus Sauer

    (University of Würzburg, Biocenter, Am Hubland)

  • Daniel Choquet

    (University of Bordeaux, CNRS, Interdisciplinary Institute for Neuroscience, IINS, UMR 5297
    University of Bordeaux, CNRS, INSERM, Bordeaux Imaging Center, BIC, UMS 3420, US 4)

Abstract

Progress in biological imaging is intrinsically linked to advances in labeling methods. The explosion in the development of high-resolution and super-resolution imaging calls for new approaches to label targets with small probes. These should allow to faithfully report the localization of the target within the imaging resolution – typically nowadays a few nanometers - and allow access to any epitope of the target, in the native cellular and tissue environment. We report here the development of a complete labeling and imaging pipeline using genetic code expansion and non-canonical amino acids in neurons that allows to fluorescently label masked epitopes in target transmembrane proteins in live neurons, both in dissociated culture and organotypic brain slices. This allows us to image the differential localization of two AMPA receptor (AMPAR) auxiliary subunits of the transmembrane AMPAR regulatory protein family in complex with their partner with a variety of methods including widefield, confocal, and dSTORM super-resolution microscopy.

Suggested Citation

  • Diogo Bessa-Neto & Gerti Beliu & Alexander Kuhlemann & Valeria Pecoraro & Sören Doose & Natacha Retailleau & Nicolas Chevrier & David Perrais & Markus Sauer & Daniel Choquet, 2021. "Bioorthogonal labeling of transmembrane proteins with non-canonical amino acids unveils masked epitopes in live neurons," Nature Communications, Nature, vol. 12(1), pages 1-16, December.
    • Handle: RePEc:nat:natcom:v:12:y:2021:i:1:d:10.1038_s41467-021-27025-w
    DOI: 10.1038/s41467-021-27025-w
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    References listed on IDEAS

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