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An Activity‐Based Sensing Approach for the Detection of Cyclooxygenase‐2 in Live Cells
Author(s) -
Yadav Anuj K.,
Reinhardt Christopher J.,
Arango Andres S.,
Huff Hannah C.,
Dong Liang,
Malkowski Michael G.,
Das Aditi,
Tajkhorshid Emad,
Chan Jefferson
Publication year - 2020
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.201914845
Subject(s) - cyclooxygenase , flow cytometry , natural product , confocal microscopy , docking (animal) , intracellular , substrate (aquarium) , peroxidase , chemistry , microbiology and biotechnology , enzyme , computational biology , biology , biochemistry , medicine , ecology , nursing
Cyclooxygenase‐2 (COX‐2) overexpression is prominent in inflammatory diseases, neurodegenerative disorders, and cancer. Directly monitoring COX‐2 activity within its native environment poses an exciting approach to account for and illuminate the effect of the local environments on protein activity. Herein, we report the development of CoxFluor, the first activity‐based sensing approach for monitoring COX‐2 within live cells with confocal microscopy and flow cytometry. CoxFluor strategically links a natural substrate with a dye precursor to engage both the cyclooxygenase and peroxidase activities of COX‐2. This catalyzes the release of resorufin and the natural product, as supported by molecular dynamics and ensemble docking. CoxFluor enabled the detection of oxygen‐dependent changes in COX‐2 activity that are independent of protein expression within live macrophage cells.