An elastase activity reporter for Electronic Paramagnetic Resonance (EPR) and Overhauser-enhanced Magnetic Resonance Imaging (OMRI) as a line-shifting nitroxide - Aix-Marseille Université Accéder directement au contenu
Article Dans Une Revue Free Radical Biology and Medicine Année : 2018

An elastase activity reporter for Electronic Paramagnetic Resonance (EPR) and Overhauser-enhanced Magnetic Resonance Imaging (OMRI) as a line-shifting nitroxide

Résumé

Pulmonary inflammatory diseases are a major burden worldwide. They have in common an influx of neutrophils. Neutrophils secrete unchecked proteases at inflammation sites consequently leading to a protease/inhibitor imbalance. Among these proteases, neutrophil elastase is responsible for the degradation of the lung structure via elastin fragmentation. Therefore, monitoring the protease/inhibitor status in lungs non-invasively would be an important diagnostic tool. Herein we present the synthesis of a MeO-Suc-(Ala)(2)-Pro-Val-nitroxide, a line-shifting elastase activity probe suitable for Electron Paramagnetic Resonance spectroscopy (EPR) and Overhauser-enhanced Magnetic Resonance Imaging (OMRI). It is a fast and sensitive neutrophil elastase substrate with K-m = 15 +/- 2.9 mu M, k(cat)/K-m = 930,000 s(-1) M-1 and K-m = 25 +/- 5.4 mu M, k(cat)/K-m = 640,000 s(-1) M-1 for the R and S isomers, respectively. These properties are suitable to detect accurately concen trations of neutrophil elastase as low as 1 nM. The substrate was assessed with broncho-alveolar lavages samples derived from a mouse model of Pseudomonas pneumonia. Using EPR spectroscopy we observed a clear-cut difference between wild type animals and animals deficient in neutrophil elastase or deprived of neutrophil Elastase, Cathepsin G and Proteinase 3 or non-infected animals. These results provide new preclinical ex vivo and in vivo diagnostic methods. They can lead to clinical methods to promote in time lung protection.

Domaines

Chimie
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Dates et versions

hal-02091886 , version 1 (07-04-2019)

Identifiants

Citer

Natacha Jugniot, Indranil Duttagupta, Angelique Rivot, Philippe Massot, Colleen Cardiet, et al.. An elastase activity reporter for Electronic Paramagnetic Resonance (EPR) and Overhauser-enhanced Magnetic Resonance Imaging (OMRI) as a line-shifting nitroxide. Free Radical Biology and Medicine, 2018, 126, pp.101--112. ⟨10.1016/j.freeradbiomed.2018.08.006⟩. ⟨hal-02091886⟩
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