Mitochondria isolated in nearly isotonic KCl buffer: focus on cardiolipin and organelle morphology.
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.
Corcelli
Angela
A
Department of Medical Biochemistry, Biology and Physics, University of Bari, Bari, Italy. a.corcelli@biologia.uniba.it
Saponetti
Matilde Sublimi
MS
Zaccagnino
Patrizia
P
Lopalco
Patrizia
P
Mastrodonato
Maria
M
Liquori
Giuseppa E
GE
Lorusso
Michele
M
D000818:Animals; D002021:Buffers; D002308:Cardiolipins / Q000378:metabolism*; D019069:Cell Respiration / Q000187:drug effects; D002855:Chromatography, Thin Layer; D003579:Cytochrome Reductases / Q000378:metabolism; D006418:Heme / Q000378:metabolism; D007552:Isotonic Solutions / Q000494:pharmacology*; D008055:Lipids / Q000032:analysis; D008930:Mitochondria, Liver / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism* / Q000648:ultrastructure; D009249:NADP / Q000378:metabolism; D053140:Organelle Shape / Q000187:drug effects*; D010101:Oxygen Consumption / Q000187:drug effects; D011189:Potassium Chloride / Q000494:pharmacology*; D051381:Rats; D021241:Spectrometry, Mass, Electrospray Ionization
D002021:Buffers; D002308:Cardiolipins; D007552:Isotonic Solutions; D008055:Lipids; D006418:Heme; D009249:NADP; D011189:Potassium Chloride; D003579:Cytochrome Reductases