Sep 10, 1976

Oxidative phosphorylation in right-side-out membrane vesicles from Escherichia coli

The Journal of Biological Chemistry
T Tsuchiya

Abstract

Oxidative phosphorylation in Escherichia coli membrane vesicles with a right-side-out orientation and loaded with ADP was investigated. Substrates of the electron transport chain could energize the phosphorylation of ADP, with the order of effectiveness being D-lactate greater than reduced phenazinemethosulfate greater than succinate greater than reduced nicotinamide adenine dinucleotide. Inhibitors of D-lactate oxidation, proton conductors, and inhibitor of the Mg2+ATPase (EC 3.6.1.3) all inhibited oxidative phosphorylation when coupled to D-lactate oxidation. ATP synthesis was absent in membrane vesicles prepared from a mutant strain lacking the Mg2+ATPase. Valinomycin or nigericin partially inhibited oxidative phosphorylation in the presence of potassium. Valinomycin plus nigericin completely inhibited ATP synthesis. The effect of various agents on the respiration-dependent establishment of a transmembrane pH gradient was also examined. NaCN and carbonyl cyanide p-trifluoromethoxyphenylhydrazone inhibited the establishment of a pH gradient while dicyclohexylcarbodiimide had no effect. These results are in good agreement with a chemiosmotic model for oxidative phosphorylation.

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Mentioned in this Paper

Dinitrophenols
Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone
Ethanol
Alkalescens-Dispar Group
Arsenates
Oxidative Phosphorylation
Hydrogen-Ion Concentration
Respiratory Chain
Plasma Membrane
Dicyclohexylcarbodiimide

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