A fluorescence probe of energy-dependent structure changes in fragmented membranes

Abstract

The reaction of the fluorochrome, 8-anilino-1-naphthalene-sulfonic acid (ANS), with fragmented membranes from beef heart mitochondria has been studied. ANS fluorescence is found to be enhanced 25-fold on binding to the membrane fragments in the absence of energy conservation, and this enhancement is increased to 35-fold in the membrane energized by substrate plux oxygen. The fluorescence of bound ANS depends upon the energy state of the membrane fragments, as indicated by the effects of ATP, substrates of the respiratory chain, oligomycin, and uncouplers. It is concluded that the changes of ANS fluorescence indicate structural changes of the mitochondrial membrane associated with energy conservation. The time course of energization is readily followed by ANS, and has a half-time of two seconds at 26 degrees .