Abstract
Publisher Summary This chapter describes a procedure to obtain a preparation of H + transporting ATP synthase from pig heart mitochondria as pure and as active for ATP synthesis as possible. The basic approach of the procedure is to extract the H + -transporting ATP synthase from the mitochondrial inner membrane after elimination of as many contaminating proteins as possible from either face of the membrane. The rate of ATP synthesis is estimated by the ATP-P i exchange based on the incorporation of 32 P i into the terminal phosphoryl group of ATP, modified by the addition of excess ADP to obtain linear rates of ATP synthesis even in preparations with low rates of ATPase activity. The vesicular preparation of H + -transporting ATP synthase is non-permeant and inverted. It yields a very efficient enzyme with high ATP synthase activity. It is a good model to study the properties of H + -transporting ATP synthase.
Published Version
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