线粒体及其亚细胞器区室和膜的分离

Isolation of Mitochondria, Their Sub-Organellar Compartments, and Membranes.

作者信息

Duncan Owen, Millar A Harvey, Taylor Nicolas L

机构信息

The ARC Centre of Excellence in Plant Energy Biology, The University of Western Australia, M316, 35 Stirling Highway, Perth, WA, 6009, Australia.

The ARC Centre of Excellence in Plant Energy Biology, The University of Western Australia, M316, 35 Stirling Highway, Crawley, Perth, WA, 6009, Australia.

出版信息

Methods Mol Biol. 2017;1511:83-96. doi: 10.1007/978-1-4939-6533-5_7.

DOI:10.1007/978-1-4939-6533-5_7

PMID:27730604

Abstract

Mitochondria are the sites of a diverse set of essential biochemical processes in plants. In order to facilitate the analysis of these functions, this chapter presents protocols for the isolation of intact mitochondria from a range of plant tissues as well two workflows for fractionation into their four subcompartments; the inner and outer membranes and the two aqueous compartments, the inter membrane space and matrix. Protocols for the assessment of mitochondrial integrity and purity through enzymatic function and suggestions of commercially available compartment marker antibodies are provided.

摘要

线粒体是植物中一系列重要生化过程的发生场所。为了便于分析这些功能，本章介绍了从多种植物组织中分离完整线粒体的方法，以及将线粒体分离为四个亚组分（内膜、外膜以及两个水相组分，即膜间隙和基质）的两种工作流程。文中还提供了通过酶功能评估线粒体完整性和纯度的方法，以及市售亚组分标记抗体的相关建议。