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

ABSTRACT: 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 successfu

SUBMITTER: Anny CA 

PROVIDER: S-EPMC12781144 | biostudies-literature | 2026 Jan

REPOSITORIES: biostudies-literature