7. The electron transport chain is relatively inaccessible to experimental manip

Question

7. The electron transport chain is relatively inaccessible to experimental manipulation in intact mitochondria. After disrupting mitochondria with ultrasound, it is possible to isolate functional sub-mitochondrial particles, which consist of broken cristae that have resealed inside out into small closed vesicles. In these vesicles the components that originally faced the matrix are now exposed to the surrounding medium. How could such an arrangement aid in the study of electron transport and ATP synthesis?

Explanation / Answer

So, once you get these particles, you examine them under an electron microscope using negative staining (BAckground is stained instead of the particles). What you see is a nobel prize winning entity - The Fo-F1 ATPase , in the form of spheres on the outer surface attached by stalks. These are seen on the outer side as the membrane reverses, i.e. they are actually on the inner membrane of the INTACT mitochondria. This gives a "Cell-free" system to study the Electron Transfer Chain and get the purified proteins in a functional state. The reaction is a couple between reducing equivalents (NADH, ADP or Pi) getting oxidised by transferring electrons to O2, and the coupling of it to synthesis of ATP via the reaction ADP + Pi -> ATP.

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