Treffer: Simultaneous Visualization of Distinct Posttranslational Modification States of β-Catenin Using Genetic Code Expansion and Click Chemistry.

Title:
Simultaneous Visualization of Distinct Posttranslational Modification States of β-Catenin Using Genetic Code Expansion and Click Chemistry.
Authors:
Anny CA; Univ. Lille, CNRS, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, F-59000, Lille, France., Nouaille S; TBI, Université de Toulouse, CNRS, INRAE, INSA, F-31077, Toulouse, France., Huvent I; Univ. Lille, CNRS, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, F-59000, Lille, France., Spriet C; Univ. Lille, CNRS, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, F-59000, Lille, France.; Univ. Lille, CNRS, Inserm, CHU Lille, Institut Pasteur de Lille, US 41 - UAR 2014 - PLBS, F-59000, Lille, France., Schulz C; Univ. Lille, CNRS, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, F-59000, Lille, France., Bray F; University of Lille, CNRS, UAR 3290-MSAP-Miniaturisation pour la Synthèse, l'Analyze et la Protéomique, F-59000, Lille, France., Fauré R; TBI, Université de Toulouse, CNRS, INRAE, INSA, F-31077, Toulouse, France., Lefebvre T; Univ. Lille, CNRS, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, F-59000, Lille, France., Biot C; Univ. Lille, CNRS, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, F-59000, Lille, France.
Source:
Chembiochem : a European journal of chemical biology [Chembiochem] 2026 Jan; Vol. 27 (1), pp. e202500547. Date of Electronic Publication: 2025 Dec 12.
Publication Type:
Journal Article
Language:
English
Journal Info:
Publisher: Wiley-VCH Verlag Country of Publication: Germany NLM ID: 100937360 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1439-7633 (Electronic) Linking ISSN: 14394227 NLM ISO Abbreviation: Chembiochem Subsets: MEDLINE
Imprint Name(s):
Original Publication: Weinheim, Germany : Wiley-VCH Verlag, c2000-
MeSH Terms:
beta Catenin*/genetics , beta Catenin*/metabolism , beta Catenin*/chemistry , Click Chemistry* , Protein Processing, Post-Translational* , Genetic Code*, Humans ; HeLa Cells ; Selenocysteine/analogs & derivatives ; Selenocysteine/chemistry ; Green Fluorescent Proteins/genetics ; Green Fluorescent Proteins/metabolism
References:
Database (Oxford). 2021 Apr 7;2021:. (PMID: 33826699)
MedComm (2020). 2023 May 02;4(3):e261. (PMID: 37143582)
Annu Rev Biophys Biomol Struct. 2006;35:225-49. (PMID: 16689635)
J Photochem Photobiol B. 2018 Nov;188:95-99. (PMID: 30240974)
Mol Cells. 2019 Jan 31;42(1):8-16. (PMID: 30699286)
Chembiochem. 2026 Jan;27(1):e202500547. (PMID: 41387094)
Cell. 2002 Mar 22;108(6):837-47. (PMID: 11955436)
Molecules. 2020 Oct 01;25(19):. (PMID: 33019562)
Angew Chem Int Ed Engl. 2007;46(36):6849-51. (PMID: 17685371)
Sci Rep. 2019 Apr 18;9(1):6298. (PMID: 31000738)
Nat Protoc. 2006;1(6):3001-10. (PMID: 17406561)
Anal Chem. 2017 Mar 21;89(6):3656-3663. (PMID: 28234450)
Biomolecules. 2025 Aug 03;15(8):. (PMID: 40867560)
FASEB J. 2014 Aug;28(8):3325-38. (PMID: 24744147)
J Pharm Sci. 2005 Feb;94(2):304-16. (PMID: 15570599)
Chembiochem. 2015 Dec;16(18):2571-5. (PMID: 26488919)
Glycobiology. 2023 Dec 30;33(12):1172-1181. (PMID: 37856504)
Biochemistry. 2017 Jul 11;56(27):3507-3517. (PMID: 28627871)
Grant Information:
847568 H2020 Marie
Contributed Indexing:
Keywords: click chemistry; genetic code expansion; posttranslational modifications; protein imaging; β‐catenin
Substance Nomenclature:
0 (beta Catenin)
0CH9049VIS (Selenocysteine)
147336-22-9 (Green Fluorescent Proteins)
Entry Date(s):
Date Created: 20251212 Date Completed: 20260108 Latest Revision: 20260110
Update Code:
20260110
PubMed Central ID:
PMC12781144
DOI:
10.1002/cbic.202500547
PMID:
41387094
Database:
MEDLINE

Weitere Informationen

A novel strategy based on genetic code expansion combined with click chemistry for the simultaneous visualization of distinct posttranslational modification (PTM) states of a single protein within living cells. As a model, it is focused on threonine 41 (T41) of β-catenin, a regulatory hotspot implicated in epithelial cancers and known to be phosphorylated, O-GlcNAcylated, or left unmodified. Using site-specific incorporation of the unnatural phenylselenocysteine, a β-catenin-EGFP fusion protein is engineered allowing selective installation of a S-GlcNAc moiety via oxidative elimination and thiol-Michael ligation. Additional β-catenin variants, phosphomimetic T41E-mCherry and wild-type-blue fluorescent protein fusions, are produced to represent other PTM states. All constructs are successfully introduced into Hep3B and HeLa cells by lipofection or TAT-mediated transduction. Fluorescence microscopy revealed distinct subcellular localization profiles for each PTM form. Notably, the S-GlcNAcylated β-catenin exhibited enhanced resistance to proteasomal degradation, consistent with known roles of O-GlcNAcylation in protein stability. This approach provides a versatile platform to functionally probe PTMs in a comparative, cell-based context.
(© 2025 The Author(s). ChemBioChem published by Wiley‐VCH GmbH.)