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Chapitre D'ouvrage Année : 2023

Analysis of ATG8 family members using LC3-interacting regions (LIR)-based molecular traps

Résumé

The ATG8 family of proteins regulates the autophagy process from the autophagosome maturation and cargo recruitment up to degradation. Autophagy dysfunction is involved in the development of multiple diseases. The LC3 interacting region (LIR)-based molecular traps have been designed to isolate endogenous ATG8 proteins and their interactors in order to facilitate the study of selective autophagy events. Here, we summarize protocols describing LC3 traps and sample preparation as well as adaptations for the analysis of ATG8 proteins in different biological models. This protocol was optimized to prepare affinity columns, reduce background, and improve the protein recovery to be analyzed by immunodetection with antibodies recognizing proteins of interest.
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Dates et versions

hal-03991599 , version 1 (06-11-2023)

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Grégoire Quinet, Pierre Génin, Naima Belgareh-Touzé, Oznur Ozturk, Robert Weil, et al.. Analysis of ATG8 family members using LC3-interacting regions (LIR)-based molecular traps. Manuel S. Rodriguez; Rosa Barrio. The Ubiquitin Code, 2602, Springer US, pp.191-204, 2023, Methods in Molecular Biology, 978-1-0716-2858-4. ⟨10.1007/978-1-0716-2859-1_14⟩. ⟨hal-03991599⟩
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