Events near the reaction center in O2 evolving PS II enriched thylakoid membranes: The presence of an electric field during the S2 state in a population of centers.

Flash-induced absorption changes at 515 nm observed as a function of flash number are examined in relation to the flash-induced fluorescence yields in inside-out thylakoids. After partial dissipation of the delocalized transmembrane electric field by adding gramicidin, the analysis of period 4 oscillations and of the kinetics in the 10 ms-1 s range suggest that the variation of the absorption changes at 515 nm as a function of flash number is the result of at least two processes:1) an electric field increase related to the S2 state and 2) the fact that the field generated by the water protons inside the membrane decreases when these protons are released outside the membrane. The former field correlates with the flash-induced fluorescence yield increase induced by the donor side of Photosystem II. Both measurements show similar oscillations as a function of flash number, with maxima on the 1st, 5th and 9th flash. These oscillations, after a shift of two flashes, appear to be different from those of the O2 yield observed under similar conditions. It is proposed that, in a population of centers the electric field during the S2 state reflects the presence of a stabilized positive equivalent in the protein close to the Mn complex.