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ACS Cent Sci. 2019 Mar 27;5(3):539-548. doi: 10.1021/acscentsci.8b00933. Epub 2019 Feb 19.

Cathepsin G Activity as a New Marker for Detecting Airway Inflammation by Microscopy and Flow Cytometry.

Guerra M1,2,3, Frey D4,3, Hagner M4,3, Dittrich S4,3, Paulsen M4,3, Mall MA4,3,5,6, Schultz C1,3,7.

Author information

1
Molecular Medicine Partnership Unit (MMPU), European Molecular Biology Laboratory (EMBL) and University of Heidelberg, 69117 Heidelberg, Germany.
2
Faculty of Biosciences, Collaboration for Joint Ph.D. Degree between EMBL and Heidelberg University, 69117 Heidelberg, Germany.
3
Translational Lung Research Center Heidelberg (TLRC), German Center for Lung Research (DZL), 69120 Heidelberg, Germany.
4
Department of Translational Pulmonology, University of Heidelberg, 69120 Heidelberg, Germany.
5
Department of Pediatric Pulmonology, Immunology and Intensive Care Medicine, Charité-Universitätsmedizin Berlin, 10117 Berlin, Germany.
6
Berlin Institute of Health (BIH), 10178 Berlin, Germany.
7
Department of Physiology and Pharmacology, Oregon Health & Science University, 3181 SW Sam Jackson Park Road, Portland, Oregon 97239-3098, United States.

Abstract

Muco-obstructive lung diseases feature extensive bronchiectasis due to the uncontrolled release of neutrophil serine proteases into the airways. To assess if cathepsin G (CG) is a novel key player in chronic lung inflammation, we developed membrane-bound (mSAM) and soluble (sSAM) FRET reporters. The probes quantitatively revealed elevated CG activity in samples from 46 patients. For future basic science and personalized clinical applications, we developed a rapid, highly informative, and easily translatable small-molecule FRET flow cytometry assay for monitoring protease activity including cathepsin G. We demonstrated that mSAM distinguished healthy from patient cells by FRET-based flow cytometry with excellent correlation to confocal microscopy data.

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