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Real‐Time Cellular Imaging of Protein Poly(ADP‐ribos)ylation
Author(s) -
Buntz Annette,
Wallrodt Sarah,
Gwosch Eva,
Schmalz Michael,
Beneke Sascha,
FerrandoMay Elisa,
Marx Andreas,
Zumbusch Andreas
Publication year - 2016
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201605282
Subject(s) - förster resonance energy transfer , context (archaeology) , microbiology and biotechnology , chemistry , fluorescence , biophysics , dna , in vitro , computational biology , biochemistry , biology , physics , paleontology , quantum mechanics
Poly(ADP‐ribos)ylation (PARylation) is an important posttranslational protein modification, and is involved in major cellular processes such as gene regulation and DNA repair. Its dysregulation has been linked to several diseases, including cancer. Despite its importance, methods to observe PARylation dynamics within cells are rare. By following a chemical biology approach, we developed a fluorescent NAD + analogue that proved to be a competitive building block for protein PARylation in vitro and in cells. This allowed us to directly monitor the turnover of PAR in living cells at DNA damage sites after near‐infrared (NIR) microirradiation. Additionally, covalent and noncovalent interactions of selected target proteins with PAR chains were visualized in cells by using FLIM‐FRET microscopy. Our results open up new opportunities for the study of protein PARylation in real time and in live cells, and will thus contribute to a better understanding of its significance in a cellular context.