Differential Scanning Calorimetry of Chloroplast Membranes: Identification of an Endothermic Transition Associated with the Water-Splitting Complex of Photosystem II

The structure of spinach thylakoid membranes has been investigated by sensitive differential scanning calorimetry. Six endotherms are observed between 20 and 85°C, corresponding to order-disorder transitions of different structural domains within the thylakoid membrane. In a medium of relatively high ionic strength, endothermic transitions occur at 42, 54, 65, 72, 79, and 84°C, with the 65°C transition being particularly prominent. At a lower ionic strength, transitions are centered at 44, 61, 66, 70, 78, and 83°C. The 42-44°C endothermic transition (the A transition) can be correlated with the modification of three electron-transport components or properties associated with photosystem II: (i) release of manganese from the membrane, (ii) the loss of O₂ evolution with water as a donor, and (iii) a decrease in the redox potential of the hydroquinone-reducible cytochrome b-559. Both the A transition and the ability to evolve O₂ are irreversibly lost after heating to 49°C and also after exposure to trypsin, suggesting the involvement of protein in this transition. The interpretation of these observations is that one effect of the A transition involves the thermal disruption of a protein component on the donor side of photosystem II.