An Activity-Based Sensing Approach for the Detection of Cyclooxygenase-2 in Live Cells

Anuj K. Yadav, Christopher J. Reinhardt, Andres S. Arango, Hannah C. Huff, Liang Dong, Michael G. Malkowski, Aditi Das, Emad Tajkhorshid, Jefferson Chan

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish (US)
Pages (from-to)3307-3314
Number of pages8
JournalAngewandte Chemie - International Edition
Volume59
Issue number8
DOIs
StatePublished - Feb 17 2020

    Fingerprint

Keywords

  • activity-based sensing
  • bioimaging
  • fluorescent probes
  • isoform selectivity
  • molecular dynamics

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)

Cite this