ISOLATION OF MITOCHONDRIAL COMPONENTS
- The first successful technique for separating inner and outer membrane developed by Donald Parson.
- In this procedure mitochondria are placed in a hypotonic solutions exposed to the detergent digitonion until the outer membrane ruptures, releasing the contents of the inter membrane space into solution.
- The inner membrane, outer membranes and components of inter membrane space are then separated by centrifugation.
- Isolated inner and outer membrane can be readily distinguished from each other by electron microscopy.
- Outer membrane book like empty sacs and inner membranes from vesicles called mitoplast containing matrix material within.
- Mitoplast can be further fractioned into inner membrane and matrix component by treating with detergent Lubrol.
- Lubrol distrups inner membrane into small inner membrane vesicles containing F1 particles.
- Biochemical studies of isolated fraction. provides important information regarding the location of various metabolic activities of the mitochondria.