title: Isolation of Mitochondria, Their Sub-Organellar Compartments, and Membranes.
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.
authors.affiliation: The ARC Centre of Excellence in Plant Energy Biology, The University of Western Australia, M316, 35 Stirling Highway, Perth, WA, 6009, Australia.
authors.forename: Owen
authors.initials: O
authors.lastname: Duncan
authors.affiliation: The ARC Centre of Excellence in Plant Energy Biology, The University of Western Australia, M316, 35 Stirling Highway, Crawley, Perth, WA, 6009, Australia.
authors.forename: A Harvey
authors.initials: AH
authors.lastname: Millar
authors.affiliation: 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.
authors.forename: Nicolas L
authors.initials: NL
authors.lastname: Taylor
mesh_terms: D000154:Aconitate Hydratase / Q000737:chemistry; D017360:Arabidopsis / Q000737:chemistry*; D029681:Arabidopsis Proteins / Q000737:chemistry*; D015415:Biomarkers / Q000737:chemistry; D002374:Catalase / Q000737:chemistry; D002458:Cell Fractionation / Q000295:instrumentation / Q000379:methods*; D002499:Centrifugation, Density Gradient / Q000295:instrumentation / Q000379:methods; D003470:Culture Media / Q000737:chemistry; D057075:Enzyme Assays; D005649:Fumarate Hydratase / Q000737:chemistry; D007425:Intracellular Membranes / Q000737:chemistry* / Q000648:ultrastructure; D007700:Kinetics; D008928:Mitochondria / Q000737:chemistry* / Q000648:ultrastructure; D020675:Peroxisomes / Q000737:chemistry* / Q000648:ultrastructure; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000737:chemistry; D011205:Povidone / Q000737:chemistry; D011523:Protoplasts / Q000737:chemistry* / Q000648:ultrastructure; D012822:Silicon Dioxide / Q000737:chemistry
chemical_list: D029681:Arabidopsis Proteins; D015415:Biomarkers; D003470:Culture Media; C016039:Percoll; D012822:Silicon Dioxide; D002374:Catalase; D017853:Phosphotransferases (Alcohol Group Acceptor); C022390:phosphoribulokinase; D005649:Fumarate Hydratase; D000154:Aconitate Hydratase; D011205:Povidone
keywords: Arabidopsis; Intermembrane space; Matrix; Mitochondria; Protoplast; Solanacea; Subfractionation