Primary charge separation within the structurally symmetric tetrameric Chl_2AP_AP_BChl_2B chlorophyll exciplex in...

In Photosystem I (PS I), the role of the accessory chlorophyll (Chl) molecules, Chl_2A and Chl_2B (also termed A_-1A and A_-1B), which are directly adjacent to the special pair P₇₀₀ and fork into the A- and B-branches of electron carriers, is incompletely understood. In this work, the Chl_2A and Chl_2B transient absorption ΔA₀(λ) at a time delay of 100 fs was identified by ultrafast pump-probe spectroscopy in three pairs of PS I complexes from Synechocystis sp. PCC 6803 with residues PsaA-N600 or PsaB-N582 (which ligate Chl_2B or Chl_2A through a H₂O molecule) substituted by Met, His, and Leu. The ΔA₀(λ) spectra were quantified using principal component analysis, the main component of which was interpreted as a mutation-induced shift of the equilibrium between the excited state of primary donor P₇₀₀^⁎ and the primary charge-separated state P₇₀₀⁺Chl₂^-. This equilibrium is shifted to the charge-separated state in wild-type PS I and to the excited P₇₀₀ in the PS I complexes with the substituted ligands to the Chl_2A and Chl_2B monomers. The results can be rationalized within the framework of an adiabatic model in which the P₇₀₀ is electronically coupled with the symmetrically arranged monomers Chl_2A and Chl_2B; such a structure can be considered a symmetric tetrameric exciplex Chl_2AP_AP_BChl_2B, in which the excited state (Chl_2AP_AP_BChl_2B)* is mixed with two charge-transfer states P₇₀₀⁺Chl_2A^- and P₇₀₀⁺Chl_2B^-. The electron redistribution between the two branches in favor of the A-branch apparently takes place in the picosecond time scale after reduction of the Chl_2A and Chl_2B monomers.