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Monitoring Autophagy Flux and Activity: Principles and Applications.
BioEssays ( IF 3.2 ) Pub Date : 2020-08-26 , DOI: 10.1002/bies.202000122 Takashi Ueno 1 , Masaaki Komatsu 2
BioEssays ( IF 3.2 ) Pub Date : 2020-08-26 , DOI: 10.1002/bies.202000122 Takashi Ueno 1 , Masaaki Komatsu 2
Affiliation
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Macroautophagy is a major degradation mechanism of cell components via the lysosome. Macroautophagy greatly contributes to not only cell homeostasis but also the prevention of various diseases. Because macroautophagy proceeds through multi‐step reactions, researchers often face a persistent question of how macroautophagic activity can be measured correctly. To make a straightforward determination of macroautophagic activity, diverse monitoring assays have been developed. Direct measurement of lysosome‐dependent degradation of radioisotopically labeled cell proteins has long been applied. Meanwhile, indirect monitoring procedures have been developed. In these assays, autophagosome marker proteins, microtubule‐associated proteins 1A/1B light chain 3B‐II (LC3B‐II) and gamma‐aminobutyric acid receptor‐associated protein‐II (GABARAP‐II) have been analyzed and the validity of the assays strongly depends on appropriate assessment of the fluctuation of LC3‐II and/or GABARAP‐II levels in the presence or absence of lysosomal inhibitors. This article describes these monitoring methods, paying special attention to the principles and characteristics of each procedure.
中文翻译：
监测自噬通量和活动：原理和应用。
巨自噬是细胞成分通过溶酶体降解的主要机制。巨自噬不仅有助于细胞稳态，还有助于预防各种疾病。由于巨自噬通过多步反应进行，研究人员经常面临如何正确测量巨自噬活动的持续问题。为了直接确定巨自噬活性，已经开发了多种监测方法。长期以来，直接测量放射性同位素标记的细胞蛋白的溶酶体依赖性降解。同时，还制定了间接监测程序。在这些测定中，自噬体标记蛋白，微管相关蛋白 1A/1B 轻链 3B-II (LC3B-II) 和 γ-氨基丁酸受体相关蛋白-II (GABARAP-II) 已被分析，分析的有效性在很大程度上取决于对波动的适当评估存在或不存在溶酶体抑制剂时 LC3-II 和/或 GABARAP-II 水平的变化。本文介绍了这些监控方法，特别关注每个程序的原理和特点。
更新日期：2020-10-22
中文翻译：
监测自噬通量和活动：原理和应用。
巨自噬是细胞成分通过溶酶体降解的主要机制。巨自噬不仅有助于细胞稳态，还有助于预防各种疾病。由于巨自噬通过多步反应进行，研究人员经常面临如何正确测量巨自噬活动的持续问题。为了直接确定巨自噬活性，已经开发了多种监测方法。长期以来，直接测量放射性同位素标记的细胞蛋白的溶酶体依赖性降解。同时，还制定了间接监测程序。在这些测定中，自噬体标记蛋白，微管相关蛋白 1A/1B 轻链 3B-II (LC3B-II) 和 γ-氨基丁酸受体相关蛋白-II (GABARAP-II) 已被分析，分析的有效性在很大程度上取决于对波动的适当评估存在或不存在溶酶体抑制剂时 LC3-II 和/或 GABARAP-II 水平的变化。本文介绍了这些监控方法，特别关注每个程序的原理和特点。
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