Probing the role of arginine 323 of the <scp>D1</scp> protein in photosystem <scp>II</scp> function

Abstract

The Mn₄ CaO₅ cluster of photosystem II (PSII) advances sequentially through five oxidation states (S₀ to S₄ ). Under the enzyme cycle, two water molecules are oxidized, O₂ is generated and four protons are released into the lumen. Umena et al. (2011) have proposed that, with other charged amino acids, the R323 residue of the D1 protein could contribute to regulate a proton egress pathway from the Mn₄ CaO₅ cluster and Tyr_Z via a proton channel identified from the 3D structure. To test this suggestion, a PsbA3/R323E site-directed mutant has been constructed and the properties of its PSII have been compared to those of the PsbA3-PSII by using EPR spectroscopy, polarography, thermoluminescence and time-resolved UV-visible absorption spectroscopy. Neither the oscillations with a period four nor the kinetics and S-state-dependent stoichiometry of the proton release were affected. However, several differences have been found: (1) the P₆₈₀ ⁺ decay in the hundreds of ns time domain was much slower in the mutant, (2) the S₂ Q_A ^- /DCMU and S₃ Q_A ^- /DCMU radiative charge recombination occurred at higher temperatures and (3) the S₀ Tyr_Z ^• , S₁ Tyr_Z ^• , S₂ Tyr_Z ^• split EPR signals induced at 4.2 K by visible light from the S₀ Tyr_Z , S₁ Tyr_Z , S₂ Tyr_Z , respectively, and the (S₂ Tyr_Z ^• )' induced by NIR illumination at 4.2 K of the S₃ Tyr_Z state differed. It is proposed that the R323 residue of the D1 protein interacts with Tyr_Z likely via the H-bond network previously proposed to be a proton channel. Therefore, rather than participating in the egress of protons to the lumen, this channel could be involved in the relaxations of the H-bonds around Tyr_Z by interacting with the bulk, thus tuning the driving force required for Tyr_Z oxidation.