Développement de nouvelles sondes photoactivables pour la caractérisation des cibles cellulaires du glutathion

Abstract : Glutathione (GSH and GSSG) takes a predominant part in apoptosis (programmed cell death), a process which is deficient in many cancers. The action of glutathione is closely related to the activity of several proteins having an affinity for the tripeptide ( Glu-Cys-Gly when reduced) and which are constituting the so-called glutathione system. A radioisotope-free photoaffinity labeling method was chosen for the identification of novel glutathione-binding proteins. We prepared news photoaffinity probe which can be connected to different biological molecules including, in our case, glutathione. Irradiation of a biological sample with this probe should lead to the formation of a covalent bond with proteins having an affinity for GSH. In a second step, a Staudinger ligation with biotin or a Click Chemistry reaction with modified biotin should allow the visualization and identification of the target proteins and the binding sites. Two detection methods are envisaged: - Chemiluminescence : the presence of biotin can be easily detected by a well known procedure using streptavidin-peroxydase and luminol - Fluorescence : coupling the probe with a fluorescent com
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Elodie-Denise Chenot. Développement de nouvelles sondes photoactivables pour la caractérisation des cibles cellulaires du glutathion. Médecine humaine et pathologie. Université Paul Verlaine - Metz, 2008. Français. ⟨NNT : 2008METZ025S⟩. ⟨tel-01752593⟩

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