Measurement of Cytochrome Kinetics in Rat Liver Mitochondria by Stopped Flow and Dual-Wavelength Spectrophotometry
Time resolution of the kinetics of the components of the respiratory chain of mitochondria permits to define a sequence of events foËowing oxygen addition to an anaerobic mitochondrial suspension. The stopped-flow analysis of mitochondrial electron carriers allows also to distinguish which of the carriers are “kinetically competent,” i.e., belong to the main oxidation-reduction pathway of the respiratory chain, from those which are located in side pathways (Fig. 1). (Chance, 1954; Chance et al., 1967). Dual-wavelength spectrophotometry (combined with the stopped-flow measurements) is necessary in order to distinguish, in the turbid suspension of the mitochondria, the specific changes in absorbance from the nonspecific changes in the background and to select precisely the redox component to be measured (Chance, 1974a). The stopped-flow employed is that designed and constructed in the workshop of the Johnson Research Foundation, University of Pennsylvania, by Professor Britton Chance (Chance, 1974b) (Fig. 2).
KeywordsRespiratory Chain Mitochondrial Suspension Flow Apparatus Nonspecific Change Stopped Flow
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