Calcium, conformational selection, and redox-active tyrosine YZ in the photosynthetic oxygen-evolving cluster

In Photosystem II (PSII), YZ (Tyr161D1) participates in radical transfer between the chlorophyll donor and the Mn₄CaO₅ cluster. Under flashing illumination, the metal cluster cycles among five S_n states, and oxygen is evolved from water. The essential YZ is transiently oxidized and reduced on each flash in a proton-coupled electron transfer (PCET) reaction. Calcium is required for function. Of reconstituted divalent ions, only strontium restores oxygen evolution. YZ is predicted to hydrogen bond to calcium-bound water and to His190D1 in PSII structures. Here, we report a vibrational spectroscopic study of YZ radical and singlet in the presence of the metal cluster. The S₂ state is trapped by illumination at 190 K; flash illumination then generates the S₂YZ radical. Using reaction-induced FTIR spectroscopy and divalent ion depletion/substitution, we identify calcium-sensitive tyrosyl radical and tyrosine singlet bands in the S₂ state. In calcium-containing PSII, two CO stretching bands are detected at 1,503 and 1,478 cm^-1 These bands are assigned to two different radical conformers in calcium-containing PSII. At pH 6.0, the 1,503-cm^-1 band shifts to 1,507 cm^-1 in strontium-containing PSII, and the band is reduced in intensity in calcium-depleted PSII. These effects are consistent with a hydrogen-bonding interaction between the calcium site and one conformer of radical YZ. Analysis of the amide I region indicates that calcium selects for a PCET reaction in a subset of the YZ conformers, which are trapped in the S₂ state. These results support the interpretation that YZ undergoes a redox-coupled conformational change, which is calcium dependent.