2016/01/12 by Franziska Doll, Annette Buntz, Anne‐Katrin Späte +6 · 1 voice
Biochemistry, Genetics and Molecular Biology · Chemistry · #Click Chemistry and Applications #Glycosylation and Glycoproteins Research #Ubiquitin and proteasome pathways
paper · doi:10.1002/anie.201503183
openalex publication_date 2016/01/12 · openalex created_date 2025/10/10 · openalex updated_date 2026/07/02
Protein glycosylation is a ubiquitous post-translational modification that is involved in the regulation of many aspects of protein function. In order to uncover the biological roles of this modification, imaging the glycosylation state of specific proteins within living cells would be of fundamental importance. To date, however, this has not been achieved. Herein, we demonstrate protein-specific detection of the glycosylation of the intracellular proteins OGT, Foxo1, p53, and Akt1 in living cells. Our generally applicable approach relies on Diels-Alder chemistry to fluorescently label intracellular carbohydrates through metabolic engineering. The target proteins are tagged with enhanced green fluorescent protein (EGFP). Förster resonance energy transfer (FRET) between the EGFP and the glycan-anchored fluorophore is detected with high contrast even in presence of a large excess of acceptor fluorophores by fluorescence lifetime imaging microscopy (FLIM).