分析酵母细胞中线粒体相关降解途径（MAD）。

4区生物学 Q3 Biochemistry, Genetics and Molecular Biology

Methods in enzymology Pub Date : 2024-01-01 Epub Date: 2024-09-20 DOI:10.1016/bs.mie.2024.09.001

Pin-Chao Liao, Liza A Pon

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引用次数: 0

摘要

线粒体对健康、疾病和衰老过程中的细胞功能至关重要。线粒体相关降解（MAD）是细胞器质量控制的一种途径，它能识别并泛素化未折叠的线粒体蛋白，利用包含 AAA-ATP 酶 Cdc48 及其辅助因子的保守分离酶复合物将其从细胞器中移除，并利用泛素-蛋白酶体系统（UPS）将其降解。在这里，我们描述了一种方法：(1) 研究候选 MAD 底物的周转和泛素化；(2) 在体外检测线粒体基质中 MAD 底物的逆转位和输出；(3) 以出芽酵母（Saccharomyces cerevisiae）为模式生物，研究 MAD 底物和 Cdc48 之间的相互作用。

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Analysis of the mitochondria-associated degradation pathway (MAD) in yeast cells.

Mitochondria are critical for cellular function in health, disease and aging. Mitochondria-associated degradation (MAD), a pathway for quality control of the organelle, recognizes and ubiquitinates unfolded mitochondrial proteins, removes them from the organelle using a conserved segregase complex, which contains an AAA-ATPase Cdc48 and its cofactors, and degrades them using the ubiquitin-proteasome system (UPS). Here, we describe an approach to (1) study the turnover and ubiquitination of candidate MAD substrates, (2) assay retrotranslocation and export of MAD substrates from the mitochondrial matrix in vitro, and (3) study interactions between MAD substrates and Cdc48 using the budding yeast, Saccharomyces cerevisiae, as a model organism.

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来源期刊

Methods in enzymology 生物-生化研究方法

CiteScore

2.90

自引率

0.00%

发文量

308

审稿时长

3-6 weeks

期刊介绍： The critically acclaimed laboratory standard for almost 50 years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Each volume is eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. Now with over 500 volumes the series contains much material still relevant today and is truly an essential publication for researchers in all fields of life sciences, including microbiology, biochemistry, cancer research and genetics-just to name a few. Five of the 2013 Nobel Laureates have edited or contributed to volumes of MIE.