Premium
Cover Picture: Protein‐Specific Imaging of O‐GlcNAcylation in Single Cells (ChemBioChem 18/2015)
Author(s) -
Lin Wei,
Gao Ling,
Chen Xing
Publication year - 2015
Publication title -
chembiochem
Language(s) - English
Resource type - Reports
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201500635
Subject(s) - förster resonance energy transfer , glycan , fluorophore , green fluorescent protein , fluorescence , fluorescence lifetime imaging microscopy , chemistry , internalization , live cell imaging , biophysics , nanotechnology , biology , biochemistry , cell , physics , glycoprotein , gene , materials science , quantum mechanics
The cover picture shows a FLIM‐FRET‐based strategy for specifically imaging the O‐GlcNAcylation state of a protein of interest (POI) inside a cell where thousands of proteins are modified with O‐GlcNAc. The POI is genetically tagged with EGFP, and the O‐GlcNAc on the POI is tagged with the TAMRA fluorophore by metabolic glycan labeling. EGFP and TAMRA form a FRET pair, and the occurrence of intramolecular FRET results in the shortening of the EGFP fluorescence lifetime ( τ ); this is detected by time‐ correlated single‐photon counting. The stringent nanometer proximity requirement of FRET enables specific imaging of the O‐GlcNAcylation state of the target protein.