The Mn cluster that catalyzes photosynthetic oxygen evolution was removed from the photosystem II (PSII) complex by treating PSII membranes with 1.0 mM NH2OH with concomitant inactivation of oxygen evolution. The cluster was reconstituted by incubating the treated membranes with 1.0 mM Mn2+, 20 mM Ca2+, 10 µM 2,6-dichlorophenolindophenol, and Cl− under illumination with continuous or flashing light to restore the oxygen-evolving capacity. This light-dependent activation (photoactivation) of oxygen evolution did not occur to a significant extent at 3 mM Cl−, but markedly accelerated at higher Cl− concentrations without showing a saturation phenomenon even at 1 M Cl−. At 10 mM Cl− only about 10% of the oxygen-evolving activity before NH2OH treatment was restored by 5-min illumination with continuous light, whereas at 600 mM Cl− about 60% of the original activity was recovered. This acceleration resulted from at least two different actions of Cl−: (1) stabilization of the intermediate state involved in the photoactivation process and (2) increase in the quantum yield of photoactivation. The stabilization of the intermediate was saturated at about 150 mM Cl−, whereas the increase in yield did not show saturation. The Cl−-induced increase in quantum yield did not involve any changes in the affinity of either Mn2+ binding or Ca2+ binding for photoactivation, but was rather ascribed to a protective effect of Cl− against inhibition of photoactivation by high concentrations of Mn2+. We also found that removal of the extrinsic 33-kDa protein from the PSII complex increased the CG requirement for photoactivation.
ASJC Scopus subject areas