Detection of Autophagy in Caenorhabditis elegans by Western Blotting Analysis of LGG-1.
Detection of Autophagy in Caenorhabditis elegans by Western Blotting Analysis of LGG-1.
复制标题
DOI：
10.1101/pdb.prot086512
复制
发表时间：
2016-02-01
影响因子：
--
通讯作者：
Meléndez A
中科院分区：
文献类型：
--
作者：
Palmisano NJ;Meléndez A
比较Atg8/LC3-I和Atg8- pe /LC3-II的数量是检测自噬诱导程度的常用方法。这是因为LC3-II数量的变化与细胞中存在的自噬体数量的变化密切相关。Atg8/LC3最初以未加工形式合成，经Atg4蛋白水解加工形成Atg8/LC3- i，然后修饰成磷脂酰乙醇胺（PE）偶联Atg8-PE/LC3- ii形式。Atg8/LC3- ii是Atg8/LC3的膜结合形式，而Atg8- pe /LC3- i是细胞质形式。Atg8-PE/LC3-II同时与自噬体的内外膜相关，因此Atg8-PE/LC3-II是唯一可靠的与完整的自噬体相关的自噬报告基因。
A common way to measure the induction of autophagy is to compare the amount of Atg8/LC3-I with that of Atg8-PE/LC3-II using western blot analysis. This is because changes in the amount of LC3-II are closely associated with changes in the number of autophagosomes present in a cell. Atg8/LC3 is initially synthesized as an unprocessed form, which is proteolytically processed by Atg4 to form Atg8/LC3-I, and then modified into the phosphatidylethanolamine (PE)-conjugated Atg8-PE/LC3-II form. Atg8/LC3-II is the membrane bound form of Atg8/LC3, while Atg8-PE/LC3-I is cytosolic. Atg8-PE/LC3-II associates with both the inner and outer membrane of the autophagosome, thus Atg8-PE/LC3-II is the only autophagy reporter that is reliably associated with completed autophagosomes.