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Abstract #3582

An Elastase Activity Reporter for EPR and OMRI as a Line-Shifting Nitroxide.

Natacha Jugniot1, Indranil Duttagupta2, Angélique Rivot1, Philippe Massot1, Colleen Cardiet1, Jean-Michel Franconi1, Pierre Voisin1, Elodie Parzy1, Eric Thiaudière1, Sylvain R.A Marque2, Abderrazzak Bentaher3, Gérard Audran2, and Philippe Mellet1

1UMR CNRS 5536, bordeaux, France, 2UMR CNRS 7273, marseille, France, 3EA7426, Faculté de Médecine Lyon Sud, Pierre Bénite, France

Neutrophils secrete proteases at inflammation sites leading to protease/inhibitor imbalance. Among them, neutrophil elastase (NE) is responsible for lung degradation via elastin fragmentation. Monitoring protease/inhibitor status non-invasively would be an important diagnostic tool. We present Meo-Suc-(Ala)2-Pro-Val-nitroxide, a line-shifting elastase activity probe for Electronic Paramagnetic Resonance (EPR) spectroscopy and Overhauser-enhanced Magnetic Resonance Imaging (OMRI). Fast and sensitive with Km= 15 ± 2.9 µM, kcat/Km= 930000 s-1.M-1, this substrate was assessed with bronchoalveolar lavage samples from Pseudomonas pneumonia mouse model. We observed a clear difference between wild type and NE deficient animals. These results can lead to new in vivo diagnostic methods and lung protection.

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