Simultaneous measurement of deltapH and electron transport in chloroplast thylakoids by 9-aminoacridine fluorescence.
Electron transport and the electrochemical proton gradient across the thylakoid membrane are two fundamental parameters of photosynthesis. A combination of the electron acceptor, ferricyanide and the DeltapH indicator, 9-aminoacridine, was used to measure simultaneously electron transport rates and DeltapH solely by changes in the fluorescence of 9-aminoacridine. This method yields values for the rate of electron transport that are comparable with those obtained by established methods. Using this method a relationship between the rate of electron transport and DeltapH at various uncoupler concentrations or light intensities was obtained. In addition, the method was used to study the effect of reducing the disulfide bridge in the gamma-subunit of the chloroplast ATP synthase on the relation of electron transport to DeltapH. When the ATP synthase is reduced and alkylated, the threshold DeltapH at which the ATP synthase becomes leaky to protons is lower compared with the oxidized enzyme. Proton flow through the enzyme at a lower DeltapH may be a key step in initiation of ATP synthesis in the reduced enzyme and may be the way by which reduction of the disulfide bridge in the gamma-subunit enables high rates of ATP synthesis at low DeltapH values.
Evron
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Department of Biology, Johns Hopkins University, 3400 North Charles Street, Baltimore, Maryland 21218, USA.
McCarty
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D000255:Adenosine Triphosphate / Q000096:biosynthesis / Q000378:metabolism*; D000585:Aminacrine; D004579:Electron Transport; D005453:Fluorescence; D005456:Fluorescent Dyes; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D006180:Proton-Translocating ATPases / Q000378:metabolism; D018724:Spinacia oleracea / Q000378:metabolism*; D020524:Thylakoids / Q000378:metabolism*
D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D000585:Aminacrine; D000255:Adenosine Triphosphate; D006180:Proton-Translocating ATPases