Exploring the Potential of β-Catenin ...
Document type :
Article dans une revue scientifique
PMID :
Permalink :
Title :
Exploring the Potential of β-Catenin O-GlcNAcylation by Using Fluorescence-Based Engineering and Imaging
Author(s) :
Kasprowicz, Angelina [Auteur]
Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 [UGSF]
Spriet, Corentin [Auteur]
Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 [UGSF]
1040364|||Plateformes Lilloises en Biologie et Santé - UAR 2014 - US 41 [PLBS] (VALID)
Terryn, Christine [Auteur]
Plateforme en Imagerie Cellulaire et Tissulaire [PICT]
Rigolot, Vincent [Auteur]
Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 [UGSF]
Hardivillé, Stéphan [Auteur]
Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 [UGSF]
Alteen, Matthew G. [Auteur]
Simon Fraser University = Université Simon Fraser [SFU.ca]
Lefebvre, Tony [Auteur]
Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 [UGSF]
Biot, Christophe [Auteur]
Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 [UGSF]
Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 [UGSF]
Spriet, Corentin [Auteur]
![refId](/themes/Mirage2//images/idref.png)
Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 [UGSF]
1040364|||Plateformes Lilloises en Biologie et Santé - UAR 2014 - US 41 [PLBS] (VALID)
Terryn, Christine [Auteur]
Plateforme en Imagerie Cellulaire et Tissulaire [PICT]
Rigolot, Vincent [Auteur]
Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 [UGSF]
Hardivillé, Stéphan [Auteur]
![refId](/themes/Mirage2//images/idref.png)
Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 [UGSF]
Alteen, Matthew G. [Auteur]
Simon Fraser University = Université Simon Fraser [SFU.ca]
Lefebvre, Tony [Auteur]
![refId](/themes/Mirage2//images/idref.png)
Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 [UGSF]
Biot, Christophe [Auteur]
![refId](/themes/Mirage2//images/idref.png)
Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 [UGSF]
Journal title :
Molecules
Volume number :
25
Pages :
4501
Publisher :
MDPI AG
Publication date :
2020-10-01
ISSN :
1420-3049
English keyword(s) :
bioorthogonal chemistry
fluorescence
glycosylation
metabolic incorporation
GFP
β-catenin
fluorescence
glycosylation
metabolic incorporation
GFP
β-catenin
HAL domain(s) :
Sciences du Vivant [q-bio]
Chimie/Chimie théorique et/ou physique
Chimie/Chimie théorique et/ou physique
English abstract : [en]
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 ...
Show more >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.Show less >
Show more >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.Show less >
Language :
Anglais
Peer reviewed article :
Oui
Audience :
Internationale
Popular science :
Non
Other project(s) or funding source(s) :
I-SITE CLICKNFRET
Administrative institution(s) :
Université de Lille
CNRS
CNRS
Collections :
Research team(s) :
Chemical Glycobiology
O-GlcNAcylation, signalisation cellulaire et cycle cellulaire
O-GlcNAcylation, signalisation cellulaire et cycle cellulaire
Research platform(s) :
Traitement de l'image et du signal pour la biologie (TISBio)
Submission date :
2020-11-27T14:24:43Z
2020-11-27T15:10:41Z
2021-01-28T14:28:45Z
2024-04-16T09:50:37Z
2020-11-27T15:10:41Z
2021-01-28T14:28:45Z
2024-04-16T09:50:37Z
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