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

Owen Duncan; A Harvey Millar; Nicolas L Taylor

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

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

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

Authors

Owen Duncan¹, A Harvey Millar², Nicolas L Taylor³

Affiliations

¹ The ARC Centre of Excellence in Plant Energy Biology, The University of Western Australia, M316, 35 Stirling Highway, Perth, WA, 6009, Australia.
² The ARC Centre of Excellence in Plant Energy Biology, The University of Western Australia, M316, 35 Stirling Highway, Crawley, Perth, WA, 6009, Australia.
³ The ARC Centre of Excellence in Plant Energy Biology and School of Chemistry and Biochemistry, The University of Western Australia, M316, 35 Stirling Highway, Crawley, Perth, WA, 6009, Australia. nicolas.taylor@uwa.edu.au.

PMID: 27730604
DOI: 10.1007/978-1-4939-6533-5_7

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.

Keywords: Arabidopsis; Intermembrane space; Matrix; Mitochondria; Protoplast; Solanacea; Subfractionation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Aconitate Hydratase / chemistry
Arabidopsis / chemistry*
Arabidopsis Proteins / chemistry*
Biomarkers / chemistry
Catalase / chemistry
Cell Fractionation / instrumentation
Cell Fractionation / methods*
Centrifugation, Density Gradient / instrumentation
Centrifugation, Density Gradient / methods
Culture Media / chemistry
Enzyme Assays
Fumarate Hydratase / chemistry
Intracellular Membranes / chemistry*
Intracellular Membranes / ultrastructure
Kinetics
Mitochondria / chemistry*
Mitochondria / ultrastructure
Peroxisomes / chemistry*
Peroxisomes / ultrastructure
Phosphotransferases (Alcohol Group Acceptor) / chemistry
Povidone / chemistry
Protoplasts / chemistry*
Protoplasts / ultrastructure
Silicon Dioxide / chemistry

Substances

Arabidopsis Proteins
Biomarkers
Culture Media
Percoll
Silicon Dioxide
Catalase
Phosphotransferases (Alcohol Group Acceptor)
phosphoribulokinase
Fumarate Hydratase
Aconitate Hydratase
Povidone