An Activity-Based Sensing Approach for the Detection of Cyclooxygenase-2 in Live Cells.
Anuj K YadavChristopher J ReinhardtAndres S ArangoHannah C HuffLiang DongMichael G MalkowskiAditi DasEmad TajkhorshidJefferson Y ChanPublished in: Angewandte Chemie (International ed. in English) (2020)
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.
Keyphrases
- molecular dynamics
- induced apoptosis
- cell cycle arrest
- flow cytometry
- oxidative stress
- adipose tissue
- density functional theory
- endoplasmic reticulum stress
- transcription factor
- protein protein
- squamous cell carcinoma
- cell death
- hydrogen peroxide
- small molecule
- molecular dynamics simulations
- deep learning
- neural network