An optimized protocol to analyze membrane protein degradation in yeast using quantitative western blot and flow cytometry

使用定量蛋白质印迹和流式细胞术分析酵母膜蛋白降解的优化方案

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作者:Felichi Mae Arines, Ming Li

Abstract

Membrane proteins (MPs) are essential in many cellular functions. To maintain proteostasis, MPs are downregulated via ubiquitination and degradation. Here, we describe an optimized protocol to analyze MP degradation using quantitative western blot and flow cytometry-based approaches. We use the degradation of Ypq1, a vacuole membrane lysine transporter, to demonstrate the protocol, which can be adapted for other organelle MPs and thus provide useful tools to study MP regulation in yeast and other model organisms. For complete details on the use and execution of this protocol, please refer to Arines et al. (2021) and Yang et al. (2020).

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