Mitochondria isolated in nearly isotonic KCl buffer: Focus on cardiolipin and organelle morphology

• Published in: Biochimica et biophysica acta Vol. 1798; no. 3; pp. 681 - 687
• Main Authors: Corcelli, Angela, Saponetti, Matilde Sublimi, Zaccagnino, Patrizia, Lopalco, Patrizia, Mastrodonato, Maria, Liquori, Giuseppa E., Lorusso, Michele
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.03.2010
• Subjects: Animals; Buffers; Cardiolipin; Cardiolipins - metabolism; Cell Respiration - drug effects; Chromatography, Thin Layer; Cristae morphology; Cytochrome Reductases - metabolism; Heme - metabolism; Isotonic Solutions - pharmacology; KCl based isolation buffer; Lipids - analysis; Mitochondria, Liver - drug effects; Mitochondria, Liver - enzymology; Mitochondria, Liver - metabolism; Mitochondria, Liver - ultrastructure; Mitochondrial lipids; Mitochondrion; NADP - metabolism; Organelle Shape - drug effects; Oxygen Consumption - drug effects; Potassium Chloride - pharmacology; Rats; Spectrometry, Mass, Electrospray Ionization; Mitochondrion; KCl based isolation buffer; Mitochondrial lipids; Cardiolipin; Cristae morphology
• ISSN: 0005-2736; 0006-3002; 1879-2642
• DOI: 10.1016/j.bbamem.2010.01.005

Rat liver mitochondria were isolated in parallel in two different isolation buffers: a standard buffer containing mannitol/sucrose and a nearly physiological KCl based solution. The two different organelle preparations were comparatively characterized by respiratory activity, heme content, microsomal and Golgi contamination, electron microscopy and lipid analyses. The substitution of saccharides with KCl in the isolation buffer does not induce the formation of mitoplasts or disruption of mitochondria. Mitochondria isolated in KCl buffer are coupled and able to maintain a stable transmembrane charge separation. A number of biochemical and functional differences between the two organelle preparations are described; in particular KCl mitochondria exhibit lower cardiolipin content and smaller intracristal compartments in comparison with the standard mitochondrial preparation.