Fractionation of rat liver mitochondrial components after short treatments with Triton X-100.

1. When rat liver mitochondria are treated with the non-ionic detergent Triton X-100, the solubilizing effects of the latter are already maximal after a few seconds. 2. We have developed a filtration technique that allows a fast separation of the solubilized and non-solubilized fractions. By means of this technique, we have been able to show differences in the solubilizing effect of Triton X-100 according to the physiological state of mitochondria. 3. In addition, the filtrate may be subjected to further fractionation by ultracentrifugation, by which two filtrate subfractions, supernatant and pellet, may be obtained. 4. Filtrates, supernatants and pellets differ from each other, and from intact mitochondria, in phospholipid and polypeptide composition, and possess a characteristic distribution of enzyme activities; they are also different from the ultrastructural point of view. 5. It is concluded that the combined techniques of short detergent treatments and fast filtration are useful in structural and functional studies of mitochondria and in the selective solubilization of mitochondrial components.