A new electrochemical gradient generator in thylakoid membranes of green algae

Using a new method of delayed luminescence digital imaging, mutants of Chlorella sorokiniana lacking the chloroplast CF0CF1 ATP synthase were isolated for the first time. Biochemical characterization of these strains indicates a lack of detectable synthesis and accumulation of the ATP synthase subunits alpha-CF1 and beta-CF1. Functional characterization indicates the presence of a permanent electrochemical gradient (DeltaMu) across the thylakoid membrane in the dark-adapted state, which is not suppressed under anaerobic conditions. Contrary to what is observed in the presence of the CF0CF1 ATP synthase, this gradient is essentially due to an electric field component DeltaPsi with no detectable DeltapH component, under both aerobic and anaerobic conditions. Neither the CF0CF1 ATP synthase nor a respiratory process can thus be responsible for a permanent gradient detected under these conditions. The previous proposal of a new ATP-dependent electrogenic pump in thylakoid membranes is supported by these results that, in addition, indicate a specificity of this new pump for ions other than protons.