Gurusamy Narasimman, Das Dipak K
Cardiovascular Research Center, University of Connecticut, School of Medicine, Farmington, CT, USA.
Methods Mol Biol. 2009;559:95-103. doi: 10.1007/978-1-60327-017-5_7.
Autophagy (Greek: Self digestion) is an intracellular process involved in removal of damaged or misfolded proteins or organelles. Damaged or misfolded proteins or organelles are first engulfed in a membraneous structure called autophagosome, and then the autophagosome fuse with lysosome to form autophagolysosome, where the contents are digested. Autophagy is a catabolic process induced during nutritional depletion via phosphatidylinositol 3 kinase pathway. Autophagy is induced in several diseases such as various cancers, heart failure, etc. When autophagy is induced, several autophagic genes are upregulated that help the formation of autophagosome. Several autophagosome specific marker proteins have been identified, among them MAP1LC3-II protein, which is cleaved from MAP1LC3-I, is specifically incorporated into the autophagosomal membrane. The formation of MAP1LC3-II can be analyzed by Western immunoblotting or immunofluorescence. Detailed methods of detection of MAP1LC3-II by Western immunoblotting and immunofluorescence are described.
自噬（希腊语：自我消化）是一种细胞内过程，参与清除受损或错误折叠的蛋白质或细胞器。受损或错误折叠的蛋白质或细胞器首先被包裹在一种称为自噬体的膜结构中，然后自噬体与溶酶体融合形成自噬溶酶体，其中的内容物被消化。自噬是一种在营养缺乏时通过磷脂酰肌醇3激酶途径诱导的分解代谢过程。自噬在多种疾病中被诱导，如各种癌症、心力衰竭等。当自噬被诱导时，几个自噬基因会上调，这有助于自噬体的形成。已经鉴定出几种自噬体特异性标记蛋白，其中从微管相关蛋白1轻链3-I（MAP1LC3-I）切割而来的微管相关蛋白1轻链3-II（MAP1LC3-II）蛋白会特异性地整合到自噬体膜中。MAP1LC3-II的形成可以通过蛋白质免疫印迹法或免疫荧光法进行分析。本文描述了通过蛋白质免疫印迹法和免疫荧光法检测MAP1LC3-II的详细方法。
