Cation control of chlorophyll a fluorescence yield in chloroplasts. Location of cation sensitive sites

We have compared the effect of various cations on the fluorescence yield of chloroplasts under several different conditions in vitro. 1. In the absence of a high energy state but in the presence of low (10 mM) concentrations of monovalent cations, divalent metal cations increase steady state chlorophyll a fluorescence yield in a manner which does not involve transport of these cations across the thylakoid membranes. 2. The kinetics of this cation-induced fluorescence rise are relatively slow, and seem to reflect cation binding (or subsequent conformational changes) to sites on the outer surface of the thylakoid membrane. 3. In the absence of monovalent cations, the apparent binding constant for Mg2+ to sites on the outer side of the membrane is low. Addition of low concentrations of monovalent cations (10 mM) competitively inhibits divalent cation binding. 4. Control of fluorescence yield is also exerted by the high energy state, and seems to involve proton/metal cation exchange at sites on the inner side of the thylakoid. 5. When isolated chloroplasts are washed and resuspended in a medium containing no added cations, the initial fluorescence level is high, but is quenched on addition of monovalent cations, sodium EDTA being much more effective than sodium chloride. It is argued that when isolated under these conditions, chloroplasts retain sufficient divalent cations to saturate external negative sites, and that the fluorescence lowering is due to their removal. 6. Some other cations, such as poly(L-lysine), can displace divalent cations from their external sites in an irreversible manner, resulting in a fixed low fluorescence yield.