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Additional resources for Biomembranes Part F: Bioenergetics: Oxidative Phosphorylation
149 Very high respiratory control ratios and rates of oxidation generally greater than 1000 nmol O per minute and milligram protein are found. Recently, rates greater than 4000 nmol O per minute and milligram protein have been observed and only these are adequate to support flight. The P/O ratios are as expected. 1The mitochondria contain high AMP levels on isolation, and AMP is formed on uncoupling, in contrast to rat liver mitochondria. This is a consequence of a dual localization of adenylate kinase in the matrix as well as in the intermembrane space.
The availability of many tubes of mitochondria from one large-scale preparation emancipates the investigator from the daily tedium of isolation. Such a stock eliminates individual differences in animals and inadvertent changes in the isolation procedure. Three parameters of mitochondrial function were tested (phyosphorylation efficiency, respiratory control, and Mg 2+ control of energized swelling), and each was preserved by this method. Principle Mitochondria are fortified with bovine plasma albumin and dimethyl sulfoxide (10%, v/v), a cryoprotective substance.
124 Liver (see also, this volume, Article ) The mitochondrial suspension is prepared from starved rats in a buffered s u cr o s e- EG T A (EDTA) medium by homogenization (glass-Teflon) or sand grinding, 125 followed by routine centrifugations, 126'127 or from isolated hepatocytes, 12s or from a tissue fractionation procedure. 6 The preparation consists of mitochondria from both parenchymal and nonparenchymal cells. These mitochondria have somewhat different properties. 129 Properties. The mitochondria oxidize intermediates of the citric acid cycle, glutamate plus malate, 3-hydroxybutyrate, la0 proline, 131 and fatty acids, all with good P/O ratios and good RCR, since phosphorylation is the limiting step.