In our earlier works, we have identified rate-limiting steps in the dark-to-light transition of PSII. By measuring chlorophyll a fluorescence transients elicited by single-turnover saturating flashes (STSFs) we have shown that in diuron-treated samples an STSF generates only F1 (< Fm) fluorescence level, and to produce the maximum (Fm) level, additional excitations are required, which, however, can only be effective if sufficiently long Δτ waiting times are allowed between the excitations. Biological variations in the half-rise time (Δτ1/2) of the fluorescence increment suggest that it may be sensitive to the physicochemical environment of PSII. Here, we investigated the influence of the lipidic environment on Δτ1/2 of PSII core complexes of Thermosynechococcus vulcanus. We found that while non-native lipids had no noticeable effects, thylakoid membrane lipids considerably shortened the Δτ1/2, from ~ 1 ms to ~ 0.2 ms. The importance of the presence of native lipids was confirmed by obtaining similarly short Δτ1/2 values in the whole T. vulcanus cells and isolated pea thylakoid membranes. Minor, lipid-dependent reorganizations were also observed by steady-state and time-resolved spectroscopic measurements. These data show that the processes beyond the dark-to-light transition of PSII depend significantly on the lipid matrix of the reaction center.
The principal function of the thylakoid membrane depends on the integrity of the lipid bilayer, yet almost half of the thylakoid lipids are of non-bilayer-forming type, whose exact functions are not fully understood. Non-bilayer lipids can be extruded from the membrane in the presence of high concentrations of co-solutes. We applied 2 M sucrose to induce lipid phase separation in isolated thylakoid membranes, following consequent structural and physiological effects. Circular dichroism spectroscopy indicated significant changes in the chiral macro-arrangement of the pigment-protein complexes, which were reversed after washing out the co-solute. Similarly, merocyanine-540 fluorescence suggested reversible changes in the lipid phases. The PSII function, as tested by chlorophyll fluorescence induction transients and time-resolved fluorescence, was almost unaffected. However, the presence of sucrose dramatically increased the PSII thermostability, which can partly be explained by a direct osmolyte effect and partly by the lipid phase separation stabilizing the stacked membrane., C. Kotakis, P. Akhtar, O. Zsiros, G. Garab, P. H. Lambrev., and Obsahuje bibliografické odkazy