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The flash-induced absorbance changes at 515 nanometers has been studied in chloroplasts and in digitonin subchloroplast particles of lettuce. The effect of various conditions and uncouplers was tested on the decay kinetics of this absorbance change and on ATP formation in the presence of phenazine methosulphate, either by continuous or flash illumination. It has been found that in chloroplasts, carbonyl cyanide m-chloromethoxyphenylhydrazone and nigericin in the presence of K(+) accelerate the decay of the 515 change and inhibit ATP formation. However, under a variety of conditions the rate of decay of the 515 absorbance change was found to be unrelated to ATP formation. Preillumination, addition of valinomycin in the presence of K(+), addition of Na(+), or divalent cations accelerate the decay of the 515 absorbance change markedly but have no effect on ATP formation. Addition of phosphorylation reagents has no effect on the decay rate beyond that obtained by Mg(2+) and inorganic phosphate. NH(4)Cl, and to some extent atebrin, while inhibiting ATP formation, do not affect the decay of the 515 absorbance change.In digitonin subchloroplast particles the decay kinetics of the absorbance change resemble that of chloroplasts, but the magnitude of the change is smaller. The pH change in this preparation is reduced much more than the 515 absorbance change.According to the chemiosmotic hypothesis, the sum of DeltaE(membrane potential) and DeltapH is the driving force for ATP formation. The lack of an increase in DeltaE in digitonin subchloroplast particles, which are practically devoid of DeltapH and have a normal ATP-forming activity, is inconsistent with the chemiosmotic hypothesis.

The Relation of the 515 Nanometers Absorbance Change to Adenosine Triphosphate Formation in Chloroplasts and Digitonin Subchloroplast Particles