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Sensitive Detection of Proteopathic Seeding Activity with FRET Flow Cytometry
Published on: December 8, 2015
Cathepsin G Activity as a New Marker for Detecting Airway Inflammation by Microscopy and Flow Cytometry
Matteo Guerra1,2,3, Dario Frey4,3, Matthias Hagner4,3
1Molecular Medicine Partnership Unit (MMPU), European Molecular Biology Laboratory (EMBL) and University of Heidelberg, 69117 Heidelberg, Germany.
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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