title: Free-flow electrophoresis for purification of plant mitochondria by surface charge.
abstract: Sample purity is the key for a successful in-depth analysis of any given subcellular proteome. The suitability of free-flow electrophoresis to assist conventional, centrifugation-based techniques in the preparation of plant mitochondria from green and non-green tissue was assessed by various means, including functional assays, immunoblots, electron microscopy and differential gel electrophoresis. Results indicated a significant increase in purity of the mitochondrial samples, highlighted specific contaminants previously reported as mitochondrial proteins, and also pointed to new means for separating plastids and peroxisomes from mitochondria in plant organellar extracts by exploiting differences in surface charge. This approach has the potential to allow a deeper and more comprehensive investigation of the Arabidopsis organellar proteomes, by providing a second dimension of separation based on surface charge in addition to conventional centrifugation purification protocols relying on size and density.
authors.affiliation: ARC Centre of Excellence in Plant Energy Biology, The University of Western Australia, 35 Stirling Highway, M316, Crawley, WA 6009, Australia.
authors.forename: Holger
authors.initials: H
authors.lastname: Eubel
authors.forename: Chun Pong
authors.initials: CP
authors.lastname: Lee
authors.forename: John
authors.initials: J
authors.lastname: Kuo
authors.forename: Etienne H
authors.initials: EH
authors.lastname: Meyer
authors.forename: Nicolas L
authors.initials: NL
authors.lastname: Taylor
authors.forename: A Harvey
authors.initials: AH
authors.lastname: Millar
mesh_terms: D017360:Arabidopsis / Q000648:ultrastructure*; D029681:Arabidopsis Proteins / Q000302:isolation & purification*; D004586:Electrophoresis / Q000379:methods*; D046529:Microscopy, Electron, Transmission; D008928:Mitochondria / Q000737:chemistry / Q000502:physiology / Q000648:ultrastructure; D013347:Subcellular Fractions
chemical_list: D029681:Arabidopsis Proteins