Energy transduction in photosynthetic bacteria. VII. Inhibition of the coupling ATPase by N-ethylmaleimide related to the energized state of the membrane.

N-Ethylmaleimide, at millimolar concentrations, irreversibily inhibits photophosphorylation and ATPase activity of photosynthetic membranes from Rhodopseudomonas capsulata. The inhibitory effect of N-ethylmaleimide is evident only the membranes are preincubated with the inhibitor in the light and in the absence of phosphorylation substrates. ADP and orthophosphate (or arsenate) exert a protective effect against the inhibition if they are present during the preillumination stage. The energization of the membrane by ATP hydrolysis, measured as ATP-induced quenching of 9-aminoacridine fluorescence, also is inhibited irreversibly by N-ethylmaleimide. Uncouplers protect the ATPase from inhibition by N-ethylmaleimide at concentrations at which they inhibit photophosphorylation. The ATPase, as measured either in the dark or in the light, is also inhibited by carbonylcyanide p-trifluoromethoxypenylhydrazone in parallel with photophosphorylation. These results are interpreted as evidence that the high-energy state of the membrane induces a conformational change of the ATPase, making it sensitive to attack by N-ethylmaleimide; this conformational change might be related to the active state of the ATPase.