Flash-induced ATP synthesis in pea chloroplasts in relation to proton flux

Abstract

Chloroplasts extracted from preilluminated pea seedling leaves have an active ATP synthase. Single turnover flashes induce ATP synthesis in these chloroplasts. This synthesis has been measured by the luminescence of the luciferin-luciferase assay system and the absorbance changes at 515 nm monitored to determine the concomitant proton flux through the ATP-synthase. Variations in the ratio H + ATP are observed and can be explained by the way ATP synthase is operating. The newly synthesized ATP molecules remain bound to the enzyme, and have variable probabilities to be hydrolysed or released from the enzyme, dependent upon the energized state of the membranes. The lowest value found for the ratio, H + ATP , is 1.90 ± 0.55, which should be very close to the true value.