Effect of ionophores A23187 and nigericin on the light-induced redistribution of Mg2+, K+ and H+ across the thylakoid membrane.
Passive redistributions of Mg2+ and K+ ions across the thylakoid membranes, occurring in association with the light-driven electrogenic influx of hydrogen ions have been examined in suspensions of broken spinach chloroplasts under a variety of conditions. (i) In accord with results of Hind el al. (Proc. Natl. Acad. Sci. U.S. (1974) 71, 1484), it was found that at a low K/Mg concentration ratio in the medium, the K-efflux is negligibly small, whereas a substantial Mg-efflux is observed. The converse is true when the K/Mg concentration ratio in the medium is high. (ii) In the presence of A23187, which was found to cause approximately a 60% inhibition of the light-induced pH-gradient, a significant influx of Mg2+ was observed in the light at a high K/Mg concentration ratio. Conversely the Mg-influx was small in the presence of A23187 when the K/Mg concentration ratio in the medium was low. Under these conditions, the Mg-influx was considerably increased upon the addition of valinomycin. A23187 was found not to affect the K-efflux in the light. (iii) The light-induced K-influx observed in the presence of nigericin also was found to be dependent on the concentration ratio of the monovalent and divalent cation. Its magnitude increased upon an increase in the K/Mg ratio. The results are interpreted in terms of a simplified model in which the total passive efflux of cations, driven by the potential set by the electrogenic proton pump, is considered to be a constant fraction of the proton influx. According to this, an increase in the flux of an ion species, induced either by raising its concentration, or by increasing its permeability through the membrane, will cause a decrease in the flux of the other cations. The relevance of the results is discussed with respect to conclusions about the involvement and relative magnitudes of the passive K and Mg effluxes across the thylakoid membrane during energization of intact chloroplasts and chloroplasts in situ.
Effect of ionophores A23187 and nigericin on the light-induced redistribution of Mg2+, K+ and H+ across the thylakoid membrane. Passive redistributions of Mg2+ and K+ ions across the thylakoid membranes, occurring in association with the light-driven electrogenic influx of hydrogen ions have been examined in suspensions of broken spinach chloroplasts under a variety of conditions. (i) In accord with results of Hind el al. (Proc. Natl. Acad. Sci. U.S. (1974) 71, 1484), it was found that at a low K/Mg concentration ratio in the medium, the K-efflux is negligibly small, whereas a substantial Mg-efflux is observed. The converse is true when the K/Mg concentration ratio in the medium is high. (ii) In the presence of A23187, which was found to cause approximately a 60% inhibition of the light-induced pH-gradient, a significant influx of Mg2+ was observed in the light at a high K/Mg concentration ratio. Conversely the Mg-influx was small in the presence of A23187 when the K/Mg concentration ratio in the medium was low. Under these conditions, the Mg-influx was considerably increased upon the addition of valinomycin. A23187 was found not to affect the K-efflux in the light. (iii) The light-induced K-influx observed in the presence of nigericin also was found to be dependent on the concentration ratio of the monovalent and divalent cation. Its magnitude increased upon an increase in the K/Mg ratio. The results are interpreted in terms of a simplified model in which the total passive efflux of cations, driven by the potential set by the electrogenic proton pump, is considered to be a constant fraction of the proton influx. According to this, an increase in the flux of an ion species, induced either by raising its concentration, or by increasing its permeability through the membrane, will cause a decrease in the flux of the other cations. The relevance of the results is discussed with respect to conclusions about the involvement and relative magnitudes of the passive K and Mg effluxes across the thylakoid membrane during energization of intact chloroplasts and chloroplasts in situ.