Reconstruction of the Water-Oxidizing Complex in Manganese-Depleted Photosystem II Preparations Using Mononuclear Manganese Complexes

Abstract— The water-oxidizing complex of chloroplast photosystem II is composed of a cluster of four manganese atoms that can accumulate four oxidizing redox equivalents. Depletion of manganese from the water-oxidizing complex fully inhibits oxygen evolution. However, the complex can be reconstituted in the presence of exogenous manganese in a process called photoactivation. In the present study, mononuclear manganese complexes with ligands derived from either nitrosonaphthol and ethylenediamine (Niten) or from diaminohexane and salicylaldehyde (Salhxn) are used in photoactivation experiments. Measurements of photoinduced changes of chlorophyll fluorescence yield, thermal dissipation using photoacoustic spectroscopy, photoreduction of 2,6-dichorophenolindophenol and oxygen evolution in manganese-depleted and in reconstituted photosystem II preparations demonstrate that photoactivation is more efficient when Niten and Salhxn complexes are used instead of MnCl₂. It is inferred that the aromatic ligands facilitate the interaction of the manganese atoms with photosystem II. The addition of CaCl₂ and of the extrinsic polypeptide of 33 kDa known as the manganese-stabilizing protein during photoactivation further enhances the recovery of electron transport and oxygen evolution activities. It is proposed that mononuclear manganese complexes are able to contribute to re-constitution of the water-oxidizing complex by sequential addition of single ions similarly to the current model for assembly of the tetranuclear manganese cluster and that these complexes constitute suitable model systems to study the assembly of the water-oxidizing complex.