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{"title":"In Vitro Dissection of Autophagy","authors":"Min Zhang, Dawei Liu, Liang Ge","doi":"10.1002/cpcb.33","DOIUrl":null,"url":null,"abstract":"<p>Autophagy is an essential cellular process for bulk degradation of cytoplasmic components through the lysosome. Underlying this process is an intricate interaction between protein factors and the cell endomembrane system, leading to a gradual maturation of the autophagic membrane. This structure sequesters a portion of the cytoplasm by the formation of a double-membrane compartment called the autophagosome. The autophagosome then delivers the cargo to the lysosome to complete degradation. The molecular mechanism accounting for the generation of the autophagic membrane is a longstanding question. Here, a cell-free approach that has been established to understand the mechanism of early autophagic membrane generation is described. This system has provided insight into the membrane source of the autophagosome, the early protein-membrane associations, and the membrane remodeling that generates the autophagosomal precursors. The cell-free assay, in combination with other established approaches (e.g., cell imaging), will facilitate a deeper understanding of the mechanism of autophagy. © 2017 by John Wiley & Sons, Inc.</p>","PeriodicalId":40051,"journal":{"name":"Current Protocols in Cell Biology","volume":"77 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2018-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpcb.33","citationCount":"2","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Cell Biology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpcb.33","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
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Abstract
Autophagy is an essential cellular process for bulk degradation of cytoplasmic components through the lysosome. Underlying this process is an intricate interaction between protein factors and the cell endomembrane system, leading to a gradual maturation of the autophagic membrane. This structure sequesters a portion of the cytoplasm by the formation of a double-membrane compartment called the autophagosome. The autophagosome then delivers the cargo to the lysosome to complete degradation. The molecular mechanism accounting for the generation of the autophagic membrane is a longstanding question. Here, a cell-free approach that has been established to understand the mechanism of early autophagic membrane generation is described. This system has provided insight into the membrane source of the autophagosome, the early protein-membrane associations, and the membrane remodeling that generates the autophagosomal precursors. The cell-free assay, in combination with other established approaches (e.g., cell imaging), will facilitate a deeper understanding of the mechanism of autophagy. © 2017 by John Wiley & Sons, Inc.
自噬的体外解剖
自噬是细胞质成分通过溶酶体大量降解的重要细胞过程。这一过程的基础是蛋白质因子和细胞内膜系统之间复杂的相互作用,导致自噬膜的逐渐成熟。这种结构通过形成一种叫做自噬体的双膜隔间来隔离一部分细胞质。然后自噬体将货物运送到溶酶体完成降解。自噬膜产生的分子机制是一个长期存在的问题。本文描述了一种已经建立的无细胞方法来了解早期自噬膜生成的机制。该系统对自噬体的膜源、早期蛋白-膜结合以及产生自噬体前体的膜重塑提供了深入的了解。无细胞检测,结合其他已建立的方法(如细胞成像),将有助于更深入地了解自噬的机制。©2017 by John Wiley &儿子,Inc。
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