Monitoring mitophagy in yeast: the Om45-GFP processing assay.

Monitoring mitophagy in yeast: the Om45-GFP processing assay.
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DOI:
10.4161/auto.5.8.9854
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发表时间:
2009-11
期刊:
影响因子:
13.3
通讯作者:
Klionsky DJ
Klionsky DJ
中科院分区:
生物学1区
文献类型:
--
作者:
Kanki T;Kang D;Klionsky DJ

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巨噬(巨噬)是真核细胞中普遍存在的一种降解过程。线粒体自噬(mitophagy)是一种选择性降解线粒体的特异性自噬。线粒体自噬被认为通过消除受损线粒体对维持这些细胞器的质量起重要作用,并参与细胞分化,而功能失调的线粒体自噬与神经退行性疾病有关;然而,线粒体自噬的机制尚不清楚。为了便于分析线粒体自噬,我们最近建立了一种简单的方法来监测酵母的线粒体自噬,即Om45-GFP处理试验。Om45-GFP是线粒体外膜蛋白。在线粒体进入液泡后,Om45-GFP被降解,释放出完整形式的GFP,通过免疫印迹检测。因此,游离GFP的数量反映了线粒体自噬的水平。
Macroautophagy (hereafter autophagy) is a ubiquitous degradative process in eukaryotic cells. Mitochondria autophagy (mitophagy) is a type of specific autophagy that degrades mitochondria selectively. Mitophagy is thought to play an important role for maintaining the quality of these organelles by eliminating damaged mitochondria, and it is involved in cellular differentiation, whereas dysfunctional mitophagy is related with neurodegenerative diseases; however, the mechanism of mitophagy is poorly understood. To facilitate the analysis of mitophagy, we recently established a simple method to monitor mitophagy in yeast, the Om45-GFP processing assay. Om45-GFP is a mitochondrial outer membrane protein. Following the uptake of mitochondria into the vacuole, Om45-GFP is degraded, releasing the intact form of GFP, which is detected by immunoblotting. Therefore, the amount of free GFP reflects the level of mitophagy.
DOI: 10.1016/s1534-5807(02)00373-8
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