@article{22e1a170914843aeae6e437893483aba,
title = "An Activity-Based Sensing Approach for the Detection of Cyclooxygenase-2 in Live Cells",
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.",
keywords = "activity-based sensing, bioimaging, fluorescent probes, isoform selectivity, molecular dynamics",
author = "Yadav, {Anuj K.} and Reinhardt, {Christopher J.} and Arango, {Andres S.} and Huff, {Hannah C.} and Liang Dong and Malkowski, {Michael G.} and Aditi Das and Emad Tajkhorshid and Jefferson Chan",
note = "Funding Information: The authors acknowledge the National Institute of Health for support (R35GM133581 to J.C.; R01GM115386 to M.G.M.; P41GM104601 and R01GM123455 to E.T.; R01GM1155884 and R03DA04236502 to A.D.). This work was supported in part by the Alfred P. Sloan Fellowship (FG‐2017–8964 to J.C.) and by the American Heart Association Scientist Development Grant (15SDG25760064 to A.D). C.J.R. acknowledges the Chemistry‐Biology Interface Training Grant (T32GM070421) and Seemon Pines Graduate Fellowship for support. Simulations were performed using the National Science Foundation computing resources through an XSEDE grant (TMGCA06N060 to E.T.) and PRAC allocation grant (ACI1713784 to E.T.). Major funding for the 500 MHz Bruker CryoProbeTM was provided by the Roy J. Carver Charitable Trust (Muscatine, Iowa; Grant #15–4521) to the School of Chemical Sciences NMR Lab. The Q‐Tof Ultima mass spectrometer was purchased in part with a grant from the National Science Foundation, Division of Biological Infrastructure (DBI‐0100085). We acknowledge Prof. Elvira de Mejia (Food Science and Human Nutrition, UIUC) for providing RAW 264.7 macrophage cells. We acknowledge the Core Facilities at the Carl R. Woese Institute for Genomic Biology for access to the Zeiss LSM 700 Confocal Microscope. Additionally, we acknowledge the Roy J. Carver Biotechnology Center at UIUC and Dr. Barbara Pilas for access to the BD LSR Fortessa Flow Cytometry Analyzer and for constructive discussions about instrumentation. We also thank Prof. Robert Gennis (Department of Biochemistry, UIUC) for access to the Strathkelvin oxygen electrode, Prof. H. Rex Gaskins (Department of Animal Sciences and Pathobiology, UIUC) and Emily (Jee‐Wei) Chen (Department of Chemical and Biomolecular Engineering, UIUC) for access and assistance with the BioSpherix C‐Chamber hypoxic incubator equipped with the corresponding OxyCycler C42 sub‐chamber controller, and Daryl Meling and Amogh Kambalyal for the procurement and dissection of the bovine vesicles. Funding Information: The authors acknowledge the National Institute of Health for support (R35GM133581 to J.C.; R01GM115386 to M.G.M.; P41GM104601 and R01GM123455 to E.T.; R01GM1155884 and R03DA04236502 to A.D.). This work was supported in part by the Alfred P. Sloan Fellowship (FG-2017?8964 to J.C.) and by the American Heart Association Scientist Development Grant (15SDG25760064 to A.D). C.J.R. acknowledges the Chemistry-Biology Interface Training Grant (T32GM070421) and Seemon Pines Graduate Fellowship for support. Simulations were performed using the National Science Foundation computing resources through an XSEDE grant (TMGCA06N060 to E.T.) and PRAC allocation grant (ACI1713784 to E.T.). Major funding for the 500 MHz Bruker CryoProbeTM was provided by the Roy J. Carver Charitable Trust (Muscatine, Iowa; Grant #15?4521) to the School of Chemical Sciences NMR Lab. The Q-Tof Ultima mass spectrometer was purchased in part with a grant from the National Science Foundation, Division of Biological Infrastructure (DBI-0100085). We acknowledge Prof. Elvira de Mejia (Food Science and Human Nutrition, UIUC) for providing RAW 264.7 macrophage cells. We acknowledge the Core Facilities at the Carl R. Woese Institute for Genomic Biology for access to the Zeiss LSM 700 Confocal Microscope. Additionally, we acknowledge the Roy J. Carver Biotechnology Center at UIUC and Dr. Barbara Pilas for access to the BD LSR Fortessa Flow Cytometry Analyzer and for constructive discussions about instrumentation. We also thank Prof. Robert Gennis (Department of Biochemistry, UIUC) for access to the Strathkelvin oxygen electrode, Prof. H. Rex Gaskins (Department of Animal Sciences and Pathobiology, UIUC) and Emily (Jee-Wei) Chen (Department of Chemical and Biomolecular Engineering, UIUC) for access and assistance with the BioSpherix C-Chamber hypoxic incubator equipped with the corresponding OxyCycler C42 sub-chamber controller, and Daryl Meling and Amogh Kambalyal for the procurement and dissection of the bovine vesicles. Publisher Copyright: {\textcopyright} 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim",
year = "2020",
month = feb,
day = "17",
doi = "10.1002/anie.201914845",
language = "English (US)",
volume = "59",
pages = "3307--3314",
journal = "Angewandte Chemie - International Edition",
issn = "1433-7851",
publisher = "John Wiley & Sons, Ltd.",
number = "8",
}