Prediction and screening of selective autophagy cargo in Saccharomyces cerevisiae

自噬是真核细胞中普遍存在的一种蛋白质降解系统。在自噬过程中，细胞质成分被封闭在称为自噬体的膜隔室中。在这项研究中，我们开发了一种方法来监测完整的自噬体离体检测荧光的GFP融合的氨基肽酶I，最好的特点选择性货物的自噬体在酿酒酵母。LC-MS/MS与随后的统计分析的组合揭示了自噬体货物蛋白的列表。我们所描述的方法将有助于分析Atg 11非依赖性选择性自噬的机制和生理意义。

Macroautophagy (autophagy) is a bulk protein-degradation system ubiquitously conserved in eukaryotic cells. During autophagy, cytoplasmic components are enclosed in a membrane compartment, called an autophagosome. In this study, we developed a method for monitoring intact autophagosomes ex vivo by detecting the fluorescence of GFP-fused aminopeptidase I, the best-characterized selective cargo of autophagosomes in Saccharomyces cerevisiae. A combination of LC-MS/MS with subsequent statistical analyses revealed a list of autophagosome cargo proteins. The methods we describe will be useful for analyzing the mechanisms and physiological significance of Atg11-independent selective autophagy.