title: Assessment of Posttranslational Modifications of ATG proteins.
abstract: Posttranslational modifications (PTMs) increase the functional diversity of proteins and play a key role in many cellular processes. Macroautophagy (hereafter simply referred to as autophagy) is an evolutionarily conserved, lysosome-dependent degradation pathway. This process is finely regulated by autophagy-related (ATG) genes widely conserved among eukaryotes from yeast to mammals. Various PTMs of ATG proteins such as phosphorylation, ubiquitination, and acetylation have been theorized to play a critical role in modulating autophagic processes and activity. In this chapter, we introduce several antibody-based tools (e.g., Western blot, Simple Western™, immunofluorescence, and immunoprecipitation) that are widely used to assess the PTMs of ATG proteins in mammalian cells.
authors.affiliation: University of Pittsburgh, Pittsburgh, PA, United States; Xiangya Hospital, Central South University, Changsha, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Xie
authors.affiliation: University of Pittsburgh, Pittsburgh, PA, United States.
authors.forename: R
authors.initials: R
authors.lastname: Kang
authors.affiliation: University of Pittsburgh, Pittsburgh, PA, United States. Electronic address: tangd2@upmc.edu.
authors.forename: D
authors.initials: D
authors.lastname: Tang
mesh_terms: D000818:Animals; D000071183:Autophagy-Related Proteins / Q000032:analysis* / Q000378:metabolism*; D015153:Blotting, Western / Q000379:methods*; D002478:Cells, Cultured; D005455:Fluorescent Antibody Technique / Q000379:methods*; D006801:Humans; D047468:Immunoprecipitation / Q000379:methods*; D008322:Mammals; D011499:Protein Processing, Post-Translational
chemical_list: D000071183:Autophagy-Related Proteins
keywords: Autophagy; Autophagy-related protein; Immunofluorescence; Immunoprecipitation; Posttranslational modification; Simple Western; Western blot