Mitochondria isolated in nearly isotonic KCl buffer: Focus on cardiolipin and organelle morphology |
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Authors: | Angela Corcelli Matilde Sublimi Saponetti Patrizia Lopalco Giuseppa E Liquori |
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Institution: | a Department of Medical Biochemistry, Biology and Physics, University of Bari, Bari, Italy b IPCF CNR, Bari, Italy c Institute of Biomembrane and Bioenergetics, CNR, Bari, Italy d Department of Animal and Environmental Biology, University of Bari, Italy |
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Abstract: | 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. |
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Keywords: | Mitochondrion KCl based isolation buffer Mitochondrial lipids Cardiolipin Cristae morphology |
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