Bioorthogonal labeling of transmembrane proteins with non-canonical amino acids allows access to masked epitopes in live neurons
| Autor: | Alexander Kuhlemann, Pecoraro, Markus Sauer, Gerti Beliu, Daniel Choquet, Chevrier N, Bessa-Neto D, David Perrais, Sören Doose, Natacha Retailleau |
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| Přispěvatelé: | Centre National de la Recherche Scientifique (CNRS) |
| Rok vydání: | 2021 |
| Předmět: |
chemistry.chemical_classification
0303 health sciences Chemistry [SDV.BA]Life Sciences [q-bio]/Animal biology [SDV]Life Sciences [q-bio] Confocal Genetic code Epitope Transmembrane protein Amino acid Cell biology 03 medical and health sciences 0302 clinical medicine Non canonical Bioorthogonal chemistry Biological imaging 030217 neurology & neurosurgery 030304 developmental biology |
| Popis: | 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 primary neurons that allows to fluorescently label masked epitopes in target transmembrane proteins in live neurons, both in dissociated culture and organotypic brain slices. This allowed us to image the differential localization of two glutamate receptor auxiliary proteins in complex with their partner with a variety of methods including widefield, confocal, and dSTORM super-resolution microscopy. |
| Databáze: | OpenAIRE |
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