Exploring the Potential of β-Catenin O-GlcNAcylation by Using Fluorescence-Based Engineering and Imaging - Université de Lille Accéder directement au contenu
Article Dans Une Revue Molecules Année : 2020

Exploring the Potential of β-Catenin O-GlcNAcylation by Using Fluorescence-Based Engineering and Imaging

Résumé

Monitoring glycosylation changes within cells upon response to stimuli remains challenging because of the complexity of this large family of post-translational modifications (PTMs). We developed an original tool, enabling labeling and visualization of the cell cycle key-regulator β-catenin in its O-GlcNAcylated form, based on intramolecular Förster resonance energy transfer (FRET) technology in cells. We opted for a bioorthogonal chemical reporter strategy based on the dual-labeling of β-catenin with a green fluorescent protein (GFP) for protein sequence combined with a chemically-clicked imaging probe for PTM, resulting in a fast and easy to monitor qualitative FRET assay. We validated this technology by imaging the O-GlcNAcylation status of β-catenin in HeLa cells. The changes in O-GlcNAcylation of β-catenin were varied by perturbing global cellular O-GlcNAc levels with the inhibitors of O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA). Finally, we provided a flowchart demonstrating how this technology is transposable to any kind of glycosylation.
Fichier principal
Vignette du fichier
P20.14 molecules-25-04501 (1).pdf (18.46 Mo) Télécharger le fichier
Origine : Fichiers éditeurs autorisés sur une archive ouverte

Dates et versions

hal-03028555 , version 1 (27-11-2020)
hal-03028555 , version 2 (28-01-2021)

Licence

Paternité

Identifiants

Citer

Angelina Kasprowicz, Corentin Spriet, Christine Terryn, Vincent Rigolot, Stephan Hardivillé, et al.. Exploring the Potential of β-Catenin O-GlcNAcylation by Using Fluorescence-Based Engineering and Imaging. Molecules, 2020, 25 (19), pp.4501. ⟨10.3390/molecules25194501⟩. ⟨hal-03028555v2⟩
89 Consultations
76 Téléchargements

Altmetric

Partager

Gmail Facebook X LinkedIn More