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Initial events of light-dependent ATP synthesis in spinach subchloroplast particles.

The kinetics of 32Pi incorporation into adenine nucleotides by subchloroplast particles in the light is studied with a continuous flow apparatus allowing measurements between 3 and 200 ms. After a short lag time from 1 to 3 ms ATP synthesis proceeds with a constant rate. During the first few milliseconds a faster labelling of ADP is detected. This labelling of ADP reaches a constant level up to 1 molecule ADP labelled per molecule of coupling factor present. The labelling pattern in ATP indicates that the labelled ADP does not equilibrate with free ADP. The addition of 32Pi to a phosphorylating system during the light phase (32Pi pulse) exhibits unchanged kinetic characteristics for labelling of ATP and ADP. These results indicate a phosphorylation of AMP to ADP being an intermediate step in photophosphorylation. In experiments carried out in the dark no label is found in ATP within the time analysed. However the labelling of ADP occurs in the same way as in the light.