Methods to analyze mitochondrial protein translocation in plant mitochondria.

4区 生物学 Q3 Biochemistry, Genetics and Molecular Biology
Methods in enzymology Pub Date : 2024-01-01 Epub Date: 2024-08-24 DOI:10.1016/bs.mie.2024.07.021
Saurabh Saha, Yanqiao Zhu, James Whelan, Monika W Murcha
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引用次数: 0

Abstract

Complex processes have evolved in plants to import proteins into mitochondria. Investigating these processes in plants provides insights into the specialised machinery and pathways that have evolved to cope with; (1) the immobile nature of plants that results in exposure to environmental stresses, and (2) the more complex cell environment due to the presence of plastids, the most prevalent being chloropalst in leaves. In this chapter, we present detailed protocols for the isolation of respiratory competent, coupled mitochondria from Arabidopsis thaliana, conducting protein import assays, and analyzing protein assembly into large multi-subunit complexes. Additionally, we present straightforward protocols for examining the localization of fluorescently tagged proteins to organelles such as mitochondria through protoplast transfections.

分析植物线粒体蛋白质转位的方法。
植物将蛋白质导入线粒体的过程十分复杂。研究植物中的这些过程可以深入了解为应对以下情况而进化出的专门机制和途径:(1) 植物的不可动性导致其暴露于环境压力,(2) 由于质体的存在,细胞环境更加复杂,其中最普遍的是叶片中的叶绿体。在本章中,我们介绍了从拟南芥中分离具有呼吸能力的耦合线粒体、进行蛋白质导入测定以及分析蛋白质组装成大型多亚基复合物的详细方案。此外,我们还介绍了通过原生质体转染将荧光标记蛋白质定位到线粒体等细胞器的直接方案。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Methods in enzymology
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.
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