Symmetry breaking in photosystem I: ultrafast optical studies of variants near the accessory chlorophylls in the A- and B-branches of electron transfer cofactors

Femtosecond absorption spectroscopy of Photosystem I (PS I) complexes from the cyanobacterium Synechocystis sp. PCC 6803 was carried out on three pairs of complementary amino acid substitutions located near the second pair of chlorophyll molecules Chl_2A and Chl_2B (also termed A_-1A and A_-1B). The absorption dynamics at delays of 0.1–500 ps were analyzed by decomposition into discrete decay-associated spectra and continuously distributed exponential components. The multi-exponential deconvolution of the absorption changes revealed that the electron transfer reactions in the PsaA-N600M, PsaA-N600H, and PsaA-N600L variants near the B-branch of cofactors are similar to those of the wild type, while the PsaB-N582M, PsaB-N582H, and PsaB-N582L variants near the A-branch of cofactors cause significant alterations of the photochemical processes, making them heterogeneous and poorly described by a discrete exponential kinetic model. A redistribution of the unpaired electron between the second and the third monomers Chl_2A/Chl_2B and Chl_3A/Chl_3B was identified in the time range of 9–20 ps, and the subsequent reduction of A₁ was identified in the time range of 24–70 ps. In the PsaA-N600L and PsaB-N582H/L variants, the reduction of A₁ occurred with a decreased quantum yield of charge separation. The decreased quantum yield correlates with a slowing of the phylloquinone A₀ → A₁ reduction, but not with the initial transient spectra measured at the shortest time delay. The results support a branch competition model, where the electron is sheared between Chl_2A–Chl_3A and Chl_2B–Chl_3B cofactors before its transfer to phylloquinone in either A_1A or A_1B sites.