Visualization of an Endogenous Mitochondrial Azoreductase Activity under Normoxic Conditions Using a Naphthalimide Azo-Based Fluorogenic Probe.

Autor: Michel L; CNRS, Institut de Chimie des Substances Naturelles, Université Paris-Saclay, UPR 2301, 91198 Gif-sur-Yvette, France., Auvray M; CNRS UMR 9187, Inserm U1196 Chemistry and Modeling for the Biology of Cancer Institut Curie,Université PSL, 91400 Orsay, France.; CNRS UMR 9187, Inserm U1196 Chemistry and Modeling for the Biology of Cancer, Université Paris-Saclay, 91400 Orsay, France., Askenatzis L; CNRS, Institut de Chimie des Substances Naturelles, Université Paris-Saclay, UPR 2301, 91198 Gif-sur-Yvette, France., Badet-Denisot MA; CNRS, Institut de Chimie des Substances Naturelles, Université Paris-Saclay, UPR 2301, 91198 Gif-sur-Yvette, France., Bignon J; CNRS, Institut de Chimie des Substances Naturelles, Université Paris-Saclay, UPR 2301, 91198 Gif-sur-Yvette, France., Durand P; CNRS, Institut de Chimie des Substances Naturelles, Université Paris-Saclay, UPR 2301, 91198 Gif-sur-Yvette, France., Mahuteau-Betzer F; CNRS UMR 9187, Inserm U1196 Chemistry and Modeling for the Biology of Cancer Institut Curie,Université PSL, 91400 Orsay, France.; CNRS UMR 9187, Inserm U1196 Chemistry and Modeling for the Biology of Cancer, Université Paris-Saclay, 91400 Orsay, France., Chevalier A; CNRS, Institut de Chimie des Substances Naturelles, Université Paris-Saclay, UPR 2301, 91198 Gif-sur-Yvette, France.
Jazyk: angličtina
Zdroj: Analytical chemistry [Anal Chem] 2024 Jan 30; Vol. 96 (4), pp. 1774-1780. Date of Electronic Publication: 2024 Jan 17.
DOI: 10.1021/acs.analchem.3c05030
Abstrakt: In this paper, we demonstrate the existence of an endogenous mitochondrial azoreductase (AzoR) activity that can induce the cleavage of N═N double bonds of azobenzene compounds under normoxic conditions. To this end, 100% OFF-ON azo-based fluorogenic probes derived from 4-amino-1,8-naphthalimide fluorophores were synthesized and evaluated. The in vitro study conducted with other endogenous reducing agents of the cell, including reductases, demonstrated both the efficacy and the selectivity of the probe for AzoR. Confocal experiments with the probe revealed an AzoR activity in the mitochondria of living cells under normal oxygenation conditions, and we were able to demonstrate that this endogenous AzoR activity appears to be expressed at different levels across different cell lines. This discovery provides crucial information for our understanding of the biochemical processes occurring within the mitochondria. It thus contributes to a better understanding of its function, which is implicated in numerous pathologies.
Databáze: MEDLINE