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Exploring the Potential of β-Catenin O-GlcNAcylation by Using Fluorescence-Based Engineering and Imaging
- Source :
- Molecules, Molecules, 2020, 25 (19), pp.4501. ⟨10.3390/molecules25194501⟩, Volume 25, Issue 19, Molecules, MDPI, 2020, 25 (19), pp.4501. ⟨10.3390/molecules25194501⟩, Molecules, MDPI, 2020, Molecules, 25 (19), pp.4501. ⟨10.3390/molecules25194501⟩, Molecules, Vol 25, Iss 4501, p 4501 (2020)
- Publication Year :
- 2020
- Publisher :
- HAL CCSD, 2020.
-
Abstract
- 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 &beta<br />catenin in its O-GlcNAcylated form, based on intramolecular F&ouml<br />rster resonance energy transfer (FRET) technology in cells. We opted for a bioorthogonal chemical reporter strategy based on the dual-labeling of &beta<br />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 &beta<br />catenin in HeLa cells. The changes in O-GlcNAcylation of &beta<br />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.
- Subjects :
- glycosylation
Recombinant Fusion Proteins
[SDV]Life Sciences [q-bio]
Green Fluorescent Proteins
Optical Imaging
Oligosaccharides
bioorthogonal chemistry
fluorescence
metabolic incorporation
GFP
β-catenin
Article
Acetylglucosamine
lcsh:QD241-441
[CHIM.THEO]Chemical Sciences/Theoretical and/or physical chemistry
lcsh:Organic chemistry
Metabolic Engineering
Fluorescence Resonance Energy Transfer
[SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology
Humans
[SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology
beta Catenin
HeLa Cells
Subjects
Details
- Language :
- English
- ISSN :
- 14203049
- Database :
- OpenAIRE
- Journal :
- Molecules, Molecules, 2020, 25 (19), pp.4501. ⟨10.3390/molecules25194501⟩, Volume 25, Issue 19, Molecules, MDPI, 2020, 25 (19), pp.4501. ⟨10.3390/molecules25194501⟩, Molecules, MDPI, 2020, Molecules, 25 (19), pp.4501. ⟨10.3390/molecules25194501⟩, Molecules, Vol 25, Iss 4501, p 4501 (2020)
- Accession number :
- edsair.pmid.dedup....5f852a17dd2b27a277b5869a388ff71c
- Full Text :
- https://doi.org/10.3390/molecules25194501⟩